Current Issue

 Volume 29 (1); 2021 

Development of cost-effective and accurate allele-specific PCR for determination of M2/ANXA5 haplotype in recurrent pregnancy loss
Kai-Cheen Ang, Timofey S. Rozhdestvensky, Arseni Markoff, Nadja Bogdanova, Ewe Seng Ch’ng,
Thean-Hock Tang
APJMBB 29 (1): 1-9
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.01
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Repeated Pregnancy Loss (RPL) affects approximately 1 out of 20 pregnant women globally; it is traumatic for parents seeking parenthood with ensuing anxieties for the next pregnancy. M2/ANXA5 haplotype is a hereditary predisposition gene for thrombophilia-associated RPL; the association between M2/ANXA5 haplotype and RPL is further ascertained in a recent meta-analysis. Precision treatments have been proposed for RPL women with M2/ANXA5 haplotype. Therefore, screening for M2/ANXA5 haplotype is highly recommended as a panel of laboratory investigations for idiopathic RPL. To date, direct sequencing of PCR products is the most common method for the determination of M2/ANXA5 haplotype; this method is however tedious, expensive and time- consuming. Hereby, we demonstrate a simple and robust allele-specific PCR (AS-PCR) that detects two inherent SNPs in a single tube, which could serve as a routine genotyping tool for M2/ANXA5 haplotype. This test is rapid, only taking maximum 4 working hours to complete the analysis. Validation of the assay by 105 clinical DNA samples yields 100% concordance rate with the DNA sequencing results.
β-globin gene cluster mutations and large deletions among anaemic patients with elevated HbF level in a tertiary teaching hospital in Kelantan, Malaysia
Siti Nor Assyuhada Mat Ghani, Rozieyati Mohamed Saleh, Wan Suriana Wan Ab Rahman, Mohd Nazri Hassan, Wan Zaidah Abdullah, Maryam Azlan, Zefarina Zulkafli
APJMBB 28 (1): 10-18
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.02
Click here to download [PDF] [Supplementary Data]
Mutations in the β-globin gene cluster can lead to β-thalassaemia, δβ-thalassaemia, hereditary persistence of foetal haemoglobin (HPFH) and some of the haemoglobin variants. The clinical and haematological spectrum of thalassaemia range from benign to pathogenic conditions including severe transfusion dependent thalassaemia. Therefore, it is important to accurately diagnose β-globin gene cluster mutations to prevent thalassaemia major or intermedia offspring. The objective of this study is to detect β-globin gene cluster mutations and large deletions among anaemic patients with elevated HbF level in a tertiary teaching hospital in Kelantan, Malaysia. This study involved 144 anaemic patients with HbF level >1.0%. High-Performance Liquid Chromatography (HPLC) was used to determine the HbF and HbA2 levels. Multiplex-ARMS (ARMS)-PCR and gap-PCR were performed for those patients with high HbA2 level (>3.2%) and normal HbA2 level (≤3.2%) to detect β-globin gene cluster mutations and large deletions respectively. The majority of patients were Malays (99.3%) with mean age of 19.99 ± 1.64 years and female 61.1% predominance. Out of 87 samples tested using multiplex ARMS-PCR against eight targeted single mutation; Cd 41/42, IVS 1–5, Cd 26, Cd 17, Cd 71/72, IVS 1–1, Cd 8/9 and -28 mutations, 65 (74.7%) patients were detected which comprises of Cd 26 (56.3%), Cd 41/42 (11.5%), compound Cd 26 and Cd 41/42 (3.4%) and IVS 1–1 (3.4%). Meanwhile, for multiplex gap-PCR which detect four types of large deletions; Thai (δβ)o-thalassaemia, HPFH-6, Siriraj J and Hb Lepore, one out of 57 patients (1.8%) was found positive with Thai (δβ)o-thalassaemia. There was a significant difference between the mean of HbF level, MCV level as well as MCH level of patients with and without β-globin gene mutations and large deletions (P<0.05). This study highlighted the presence of various types of β-globin gene cluster mutations detection in establishing a definitive diagnosis among this selected group of patients for the large-scale screening of the thalassaemia gene.
Isolation and characterization Phage UPM1705 against multi-drug resistant K. pneumoniae 1705
Omar Assafiri, Adelene Ai-Lian Song, Irwan Hanish, Geok Hun Tan, Khatijah Yusoff
APJMBB 28 (1): 19-25
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.03
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The rise in in the number of drug-resistant bacteria that can resist almost all kinds of antibiotics is due to the overuse of these antibiotics (e.g., carbapenems). Thus, there is a need to find an alternative to antibiotic treatment such as the use of phages. In this study, phage UPM1705 was isolated from a polluted lake which can lyse its host Klebsiella pneumoniae ATCC BAA-1705. Based on morphological appearance from transmission electron microscopy, UPM1705 belongs to Caudovirales (Myoviridae). UPM1705 can reach a titer of 10PFU/ml based on the double-layer method. It has a burst size of 298 PFU/bacteria cell and a latent period of 80 min, a rise period of 75 min, and adsorption time of 20 min based on a one-step growth curve assay using an MOI of 0.02. It was stable from 4°C to 80°C and retained its functionality at pH between 4 to 11, with pH of 7 being the optimum pH for the phage growth. The efficiency of UPM1705 was tested via a turbidity assay at MOI of 0.02, 0.2, and 2. UPM1705 was able to clear the turbidity of the host bacteria culture at all of these three MOIs. Thus, UPM1705 has the potential to be used for phage therapy. 
Trichostatin A and Zebularine along with E-cadherin re-expression enhance tumour necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cell cycle arrest in human breast adenocarcinoma cells
Sonia How Ming Wong, Chee-Mun Fang, Hwei-San Loh, Siew Ching Ngai
APJMBB 28 (1): 26-41
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.04
Click here to download [PDF] [Supplementary Data]
Breast cancer is the leading cause of death among women in which its definite cure remains uncovered. Tumour necrosis factor-related apoptosis inducing ligand (TRAIL) is a potential anti-cancer agent due to its selective killing towards cancer cells while sparing the healthy cells. However, it is limited by the development of TRAIL resistance. With the attempt to overcome TRAIL resistance, this research embarked to study the effect of epigenetic drugs, Trichostatin A (TSA) and Zebularine (Zeb) along with E-cadherin re-expression on anti-cancer effect in human breast adenocarcinoma cells. The MDA-MB-231 re-expressed with E-cadherin (231-EGFP) was treated with TSA and Zeb before being treated with TRAIL (TZT) to compare the effect on MDA-MB-231 and MCF-7. The cell viability, cell cycle and migration assays were conducted on these cells, prior to reverse-transcription-polymerase chain reaction (RT-PCR) targeted on proliferating cell nuclear antigen (PCNA), cyclin-dependent kinase 2 (CDK2), matrix metalloproteinase 9 (MMP9). TZT induced a significant increase in G0/G1-arrested cell population and reduction in cell viability in 231-EGFP. These were verified by the suppression of PCNA and CDK2 mRNA expression. However, there was a negligible effect to reduce the cell migration of the invasive MDA-MB-231 and 231-EGFP cells in accordance with the lack of down-regulation of MMP9. In conclusion, this research shows that TSA and Zeb have sensitized breast cancer towards TRAIL treatment in 231-EGFP cells, validating the potentiality of E-cadherin as a biomarker of TRAIL treatment efficacy in the invasive breast cancer.
Preliminary study on the effect of nitrogen fertilization on growth quality of Gynura procumbens
Mohamad Fhaizal Mohamad Bukhori, Hawa Z. E. Jaafar, Ali Ghasemzadeh, Uma Rani Sinniah
APJMBB 28 (1): 42-51
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.05
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G. procumbens is an herbaceous medicinal plant. Despite the progressive reports on the potential pharmacological properties, many are overlooking at the importance of its agronomic requirements, especially in fertilization to produce high yields which have not been conclusively concluded. This study was carried out to examine the effects of nitrogen on growth quality. In this study of two factors, harvesting times and rate of nitrogen, a total of 12 combined treatments replicated three times with 108 plants were laid out according to randomized complete block design. Growth attributes were recorded in three harvests of triplicate samples to exhibit the trend for plant growth and quality, and statistically analysed. Nitrogen affected plant growth significantly (p≤0.05) with stronger effect on physiological attributes (p≤0.01). The results suggested that 0 nitrogen kg/ha at Week 12 and 90 nitrogen kg/ha at Week 4 is highly and least, respectively affected the morphology and physiology of G. procumbens. These attributes are controlled by the availability of usable nitrogen and agronomic management because nitrogen is needed in optimum quantity and must be in balance with other nutrients to achieve its maximum yield potential.
Detection of Tannerella forsythia from saliva samples in different ethnic majority groups in Sarawak
Elexson Nillian, Grace Bebey, Fatin Nabilah Ngu, Nur Diyana, Amirah Zakirah, Eddy Boli, Melvin Chung Hsien Liang
APJMBB 28 (1): 52-59
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.06
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Nowadays racial and ethnic differences in health care has become a growing concern. It is one of the critical determinant in influencing the genotype of the host in which may results in some diseases such as periodontal disease. Tannerella forsythia can be found in oral cavity and have the strongest relation in resulting on the destruction of connective tissue in periodontal disease. This research is aim to investigate the prevalence of periodontal pathogens, particularly T. forsythia in four major ethnic groups in Sarawak which may result in periodontal disease in Sarawak. This disease may due to the results from the infection of the tissue supporting the teeth. A total of (n = 40) saliva samples consist of 10 samples for each ethnic groups such as Iban, Malay, Chinese and Bidayuh were collected in Kuching and Kota Samarahan using culture-independent method. The DNA was extracted from saliva based on Phenol Chloroform Isoamyl Alcohol method. After that, 16S rRNA gene was then amplified via PCR for bacterial detection using 27 F and 1492 R primers, followed by PG-F and PG-R primers set in identifying T. forsythia. The PCR product was observed on 1.5% gel electrophoresis. As a result,  the presence of bacteria T. forsythia was found more frequently from saliva samples of ethnic in Iban (70%), followed by Malay (60%), Bidayuh (60%) and lastly Chinese (50%). The differences of demographic, certain cultural beliefs and practices might affect the oral health status. This finding show that it may help to identify the risk groups and has contributed an additional evidence for the association between ethnicity and periodontal disease.
Preliminary evaluation of potential prebiotic capacity of selected legumes and seed mucilage on the probiotic strainLactobacillus rhamnosus GG
Ka Wai Lai, Yu Hsuan How, Hasanah Mohd Ghazali, Liew Phing Pui
APJMBB 28 (1): 60-72
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.07
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The characterization of the prebiotic potential of legumes and mucilage are essential and crucial for the development of symbiotic food products. The aim of the present study was to compare and to determine the prebiotic capacity of selected legumes, namely adzuki bean, mung bean, black cowpea, and mucilages from chia seed and flaxseed on the growth of Lactobacillus rhamnosus GG. Resistance towards acid, pancreatin hydrolysis, and the prebiotic scores of the tested substances was determined based on growth promotion after 24 h of incubation. Results revealed that flaxseed mucilage was more resistant against hydrolysis by acid (1.47%) and pancreatin (2.64%) compared to the other samples (5.64 – 18.06% for acid and 10.34 – 15.57% for pancreatin). The relative prebiotic scores for flaxseed mucilage and black cowpea were 98% and 94%, respectively, which were higher than commercial prebiotics including inulin, fructooligosaccharides, and isomaltooligosaccharides. The optimum concentrations of flaxseed mucilage and black cowpea during 36 h of fermentation were 0.8% and 0.4% (w/v), respectively. The findings indicated that flaxseed mucilage was partially digested during gastrointestinal transit and it promotes the growth of the L. rhamnosus GG. The potential prebiotic capacity of flaxseed mucilage and its symbiotic relationship with L. rhamnosus GG suggests that they can be incorporated together for the development of functional foods.
Disease of consumption to extensively drug resistance tuberculosis: Race of techniques to enhance diagnosis
Jureka Mankotia, Akant Kaushal, Azhar Khan
APJMBB 28 (1): 73-81
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.1.08
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Tuberculosis is one of the most ancient infectious diseases known to mankind. Detection of the causative organism, Mycobacterium tuberculosis, has experienced a series of advances from the late 19th century through early 21st century. The hierarchy of diagnosis ranges from smear microscopy with traditional Ziehl–Neelsen staining to advanced light-emitting diode–fluorescence microscopy staining to both solid and liquid culture, and more recently, to genotypic methods like nucleic acid amplification. Each procedure has its own merits and drawbacks in terms of specificity, sensitivity, minimum detectible number of bacilli, and most crucially, turnover time. The latest diagnostic tools, like geneXpert and line probe assays, have gone a step further by detecting drug resistance. Both early and accurate diagnosis based on the drug susceptibility test implies precise and prompt initiation of treatment, thereby curtailing not only the agony of patient but also the burden tuberculosis places on the health system.



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4th International Conference on Molecular Biology & Biotechnology

in conjunction with the 27th  Scientific Meeting of the Malaysian Society of Molecular Biology & Biotechnology (MSMBB) 

Beyond 2021: Building Sustainable Science

1-3rd June2021

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