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 29 (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 29 (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 29 (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 29 (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 29 (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 29 (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 29 (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.


Volume 29 (2); 2021 

Analysis of copper removal using different seafood wastes by differential pulse voltammetry (DPV)
Joel Ashervin Zachariah, Mohd Rawa Ispal, Michelle Soo Oi Yoon
APJMBB 29 (2): 1-7
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.01
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Current methods of heavy metal removal are shown to have several drawbacks: cost of operation, generation of toxic sludge and complicated procedures in the treatment. In this study, the ability of seafood waste as a sustainable biosorbent was investigated. Three commonly consumed seafood wastes in Malaysia were chosen for this study: crab shells, prawn shells and cockle shells. This study aimed to determine the differences in copper removal between crab, prawn and cockle shells at fixed concentrations and weights of shells. Differential pulse voltammetry using a modified screen-printed electrode was used to determine the concentration of copper for each waste sample. Removal of copper was first observed at different shell weights between 20 and 100 mg and at a fixed copper concentration of 20 ppm.  Shell weights with the highest removal, 20 mg and 40 mg, were then used for further observation at concentrations between 1-20 ppm. At a weight of 20 mg, prawn shells showed the highest removal percentage of copper at 34.40% while crab shells and cockle shells showed the highest removal percentage of 31.22% and 12.78%, respectively. This was likely due to the presence of calcium carbonate and chitin as the main component in crab and prawn shells. Cockle shells showed low removal efficiency under these conditions. In conclusion, untreated crab shells and prawn shells were shown to successfully remove copper in water. 
Genetic diversity and population structure of Ethiopian faba bean (Vicia faba L.) germplasm revealed by ISSR markers
Bullo Neda, Tileye Feyissa, Kifle Dagne, Ermias Assefa
APJMBB 29 (2): 8-25
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.02
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Molecular characterization of faba bean accessions is important for future collection, conservation, and crop improvement programs. However, molecular genetic diversity of the Ethiopian faba bean accessions has not been comprehensively characterized. The present study was conducted to determine the genetic diversity and population structure of 90 faba bean landraces and six released varieties using nine inter-simple sequence repeat (ISSR) markers. The polymorphic information content (PIC), marker index (MI), resolving power (RP) and effective multiplex ratio (EMR) showed average values of 0.32, 5.87, 7.14, and 18.34 per primer, indicating high polymorphism values. The marker revealed average gene diversity of 0.26, ranging from 0.15 to 0.36. The degree of polymorphism among the landrace accessions ranged from 44.91% for the Sidama population to 72.46% for the East Hararghe population, with a mean of 57.27%. Analysis of molecular variance (AMOVA) indicated 99 % variation within and 1% variation among population, respectively. The present study revealed little to moderate population differentiation accompanied by high gene flow. Both distance-based and model-based cluster analysis distinguished seven distinct groups illustrating the conformity of results obtained from the study. Accessions from different administrative zones (AZs) of the country were distributed over most of the clusters; exhibiting high genetic diversity in collection sites. Therefore, further collection, conservation, and utilization programs would be recommended from AZs with high genetic diversity.
Anti-mycobacterial compounds derived from marine organisms: Mode of actions and potential in tuberculosis treatment
Apsara S. Babu, K. V. Bhaskara Rao
APJMBB 29 (2): 26-34
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.03
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Mycobacteria are an extremely large group of bacteria that are normal inhabitants of soil and water. It has an exceptional propensity to adjust to varying environmental conditions which has led to the success of this pathogen. Mycobacterium tuberculosis can inhabit human tissues for years without replicating, thereby resuming growth and resulting in active tuberculosis with an increased latency period compared to any other deadly disease. The existing vaccines and drugs in the market to treat tuberculosis are ineffective and pose severe side effects. Hence, the development of a novel drug with high efficacy to kill these replicating and non-replicating bacilli appears to be an attractive solution to tackle this problem. The extraction of novel biologically active compounds from marine organisms has prospective biomedical applications for future drug discovery. Tuberculosis treatment, in specific, may benefit from the identification of novel and efficient marine-based compounds. A few anti-mycobacterial compounds belonging to the class alkaloids, terpenoids, peptides, and steroids have been isolated from marine organisms and are known to have potential activity against mycobacteria. This review focuses on the currently available anti-mycobacterial compounds and quorum sensing inhibitors of marine origin along with their mode of action and their significant potential in the treatment of mycobacteria associated strains.
Screening of ligninolytic fungi for bioremediation of dyes
Seri Amelie Muliyadi, Elida Tengku Zainal Mulok, Noor Hana Hussein, Rohana Mat Nor
APJMBB 29 (2): 35-42
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.04
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Water pollution is a growing concern worldwide. One of the main causes of water pollution includes the textile industry which produces a large amount of wastewater every day. This wastewater is known to contain dyes that are recalcitrant and hard to treat. In order to solve this problem, bioremediation using ligninolytic fungi is commonly used for the ligninolytic enzymes which are able to break down the dyes. In this study, samples were collected from decaying woods and soils in the vicinity of UiTM Puncak Alam forests. A total of 20 fungal isolates were tested for ligninolytic enzyme production. Out of the 20 isolates, 13 were found to produce lignin peroxidase and manganese peroxidase, but only one produced laccase. The isolate that produced all three enzymes was used for DNA isolation and identified using amplification of the ITS region by PCR. The isolate was identified as Trichoderma asperellum, a soft rot fungal species which is renowned for its role in bioremediation as a biosorbent.
Development of a recombinant Taq DNA polymerase enzyme expressed using a synthetic gene and its comparison with a commercial enzyme
Yuliana, Uus Saepuloh, Suryani
APJMBB 29 (2): 43-50
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.05
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Taq DNA polymerase is a thermostable enzyme widely used for DNA amplification in the PCR technique. It was initially characterized and isolated from thermophilic bacteria, Thermus aquaticus. It was difficult to developed in this enzyme using a native host system. Therefore, the development of the recombinant Taq DNA polymerase expressed using a synthetic gene is important to improve production efficiency. In this study, we developed the in house Taq DNA polymerase recombinant based on a codon-optimized using E. coli expression system. We cloned 2685 bp of the Taq DNA polymerase gene in the pET151/D-TOPO vector. The gene was synthesized  and the expression was analyzed with SDS-PAGE technique which indicated with a 100.9 kDa specific target protein. The concentration and activity of this purified enzyme were 5.17 mg/mL and 4.647 U/µL, respectively. The application of this enzyme to the PCR technique showed that this enzyme could amplify the target genes from 200 bp to 3500 bp amplicons with a minimum DNA concentration template 10 ng/µL. This assumes that the in house recombinant Taq DNA polymerase based on synthetic genes is successfully expressed, purified, and was functional and comparable to the commercial Taq polymerase.
Cerium oxide nanoparticles induced physio-biochemical, neurochemical, and morphological responses in Cirrhinus mrigala during short term exposure
Kalpesh Swamy, Naveenkumar Chandrashekar, Raghunandhakumar Subramanian, Sandya Sukumaran, Sharath Chandra SP
APJMBB 29 (2): 51-61
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.06
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Cerium oxide nanoparticle (CeO2NPs) has wide applications in pharmaceutical, biomedical and chemical industries. Albeit of their uses, bioavailability followed by toxicity of CeO2NPs in fresh water fishes, are yet to be understood in detail. In this evaluation, we have synthesized, characterized and assessed the biological effects (hematology, ionoregulatory, oxidative stress, histological and glutamate indices) of CeO2NPs at different doses (2.5mg/L and 25mg/L based on 1/10th LC50) on freshwater carps Cirrhinus mrigala, for short term exposure of 96 h. Impact of CeO2NPs at low concentration (2.5mg/L) confirmed a significant decrease in hematological parameters and also affecting serum Na+, Cl-, K+ levels along with gill Na+/K+-ATPase activity. The indicated variations oxidative stress enzymes superoxide dismutase, Catalase, glutathione peroxidase with relative elevation in lipid peroxidation (LPO) (22.47±0.198) compared to control groups. CeO2NPs at high concentration (25mg/L) revealed the alterations in neurotransmitter glutamate levels compared to control groups. Rise in glucose and decrease in plasma protein levels in response to both the concentrations was noted. Microscopic observations confirmed the tissue damages and alterations in gill architecture. By integrating all results obtained by short term exposure of juvenile carps to CeO2NPs at different doses, we reported nanoparticles have considerable deleterious effects on physiological and morphological condition of fishes.
Mutations in two neuroblastoma rat sarcoma oncogenes are associated with progression of haematologic malignancies in Nigeria
Nneoma Confidence JeanStephanie Anyanwu, Ahmed Babangida Suleiman, Elijah Ekah Ella, Haruna Makanjuola Kazeem, Maryam Aminu
APJMBB 29 (2): 62-74
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.07
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Although mutation in the RAS genes has become important in the evaluation of haematologic malignancies worldwide, developing countries like Nigeria are yet to integrate it as a diagnostic tool and prognostic indicator for improved therapy. This study determined mutations in codons 12 and 13 of NRAS gene in blood donors and haematologic malignant individuals using multiplex (AS-PCR) and Sanger sequencing, thus highlighting the mutations as helpful diagnostic and prognostic tool. AS-PCR was used to selectively amplify mutation-specific synthetic oligonucleotides from the cfDNA of 100 study participants (50 cancer patients and 50 blood donors). Percentage mutation of 31.0% was seen in NRAS_G12D gene while NRAS_G13C had 17.0%. Twenty nine (29.0%) of the NRAS_G12D mutations were found in haematopoietic malignant patients and 2.0% were found in blood donors, while 15.0% of the NRAS_G13C were found in the malignant patients, confirming the occurrence of NRAS gene mutations in haematologic cancers and predominance of the G-A transition. The highest rate of mutation was observed in leukaemia patients, having a significant association with codon 13 (p = 0.042). Stages 3 and 2 cancers each had the highest mutation rates of NRAS_G12D and NRAS_G13C, revealing possible link between these mutations and susceptibility and progression of haematologic malignancies, which is higher in leukaemia. Further NRAS mutation studies and its role in other cancers are advocated, especially targeted towards ameliorating diagnosis and prognostic therapy. Challenges related to diagnosis and management of haematologic cancer continue to persist in developing countries like Nigeria. Thus, there is a need to go beyond studying the incidence and distribution pattern of these malignancies to capturing immunogenetic parameters of affected individuals.
Analysis of allele frequencies of the selected 15 autosomal STR markers in Tikrit population – Iraq with comparison to Middle Eastern, African, and Europeans
Maan Hasan Salih, Akeel Hussain Ali Al-Assie, Majeed Arsheed Sabbah
APJMBB 29 (2): 75-84
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.08
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Short tandem repeats (STRs) have been recommended as the highest polymorphic loci among the humana DNA regions. Therefore, STRs are agreeable to many genetic fields like forensic, population genetics and anthropological studies. The main aim of this research is to evaluate the autosomal STRs in Tikrit city-Iraq, to expand the human genetics database and forensic genetics analysis. The DNA database was obtained from 306 unrelated volunteers from native Tikrit population-Iraq, using 15 autosomal STR loci. The current study determined the allele frequencies in the Tikrit population and then compared them with other national Iraqi populations as well as with populations in the Middle East, Africa, and Europe. The highest level of heterozygosity was observed in D8S1179 and TH01 loci (0.797), while the less level was shown by CSF1PO (0.48). The departure from HWE Equilibrium was recorded in only 3 STR loci from a total of 15 loci analyzed (p<0.003). The Combined Match Probability (CMP) for 15 autosomal STR was 1 in 7.89208×10-19 and the Combined Discrimination Power (CDP) was 0.9999999997. The discrimination power (DP) was especially high in D2S1338, D18S51, D19S433 and D21S11. Based on the results observed in a Dendrogram, Tikrit population was clustered with other populations, likely reflecting the historical and geographical factors. D2S1338, D18S51, D19S433 and D21S11 markers were recognized as suitable for forensic genetics analysis in Tikrit population. Also, the 15 STRs markers provide information for the studies of genetic distances between the current study and other included populations to be compared with this study.
Isolation screening and molecular characterization of zinc solubilizing bacteria and their effect on the growth of wheat (Triticum aestivum)
Vandana Nandal, Manu Solanki
APJMBB 29 (2): 85-97
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.09
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Wheat (Triticum aestivum) is a major cereal crop grown worldwide. Most of the world population depends on wheat for their nutrient requirement. Zinc (Zn) is one of the most crucial elements required for the development of wheat plant. It is one of the micronutrients required in many biochemical cycles. It has been found that the concentration of Zn is below the required level in the soil and hence it remains deficient in the crops. To ameliorate the deficit, chemical fertilizers are added in the soil, where as biofertilizers are preferred over chemicals in sustainable agriculture. The paper describes the isolation, screening and molecular characterization of the zinc solubilizing bacteria (ZSB) to improve plant growth. A total of 100 soil samples were collected from the rhizospheric soil of wheat plants. ZSB were isolated by dilution plating on Bunt and Rovira media. The 50 isolates were selected and screened for their Zn solubilization. The zinc tolerance of all the isolates varied from 0.5% to 2% of insoluble Zn. Based on the Zn tolerance ability, 15 bacterial isolates were screened for Phosphate solubilization and further analyzed for the synthesis of IAA, NH3, siderophore production and chitinase activity. The three isolates were selected on the basis of the plant growth promoting characteristics for molecular characterization and were found to be homologous to Bacillus cereus, Pseudomonas aeruginosa and Bacillus tropicus. This study documented the establishment and survival of ZSB in the wheat rhizosphere and enhanced plant productivity, thus indicating the potential of isolates as commercial biofertilizers.
Expression of telomeric repeat binding factor 2 (TERF2) in childhood acute lymphoblastic leukemia
Lelamekala Vengidasan, Choong Soo Sin, Chen Kok Siong, Kamariah Ibrahim
APJMBB 29 (2): 98-106
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.2.10
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Acute lymphoblastic leukemia (ALL) is the most frequent cancer among children. Regardless of the advances in disease treatments, approximately 10-20 % of childhood ALL (cALL) have an incidence of relapse. Therefore, identification ofadditional prognostic variables is essential to provide specific therapeutic intervention for each patient. TERF2 is one of the main components of the shelterin complex (telosome) that plays a crucial role in the protective activity of telomeres. This research aims to investigate the expression level of TERF2 and its potential as a prognostic marker in cALL patients. 88 bone marrow samples and 6 peripheral blood were used to isolated cDNA samples. Real time PCR were used to study the gene expression of TERF2 in cALL. Results were standardized using B2M transcripts as an internal control. Relative quantification of the gene expression was calculated by using the delta-delta Ct method. TERF2 was up-regulated significantly in cALL patients compared to control samples of which p-value=0.002859, (p<0.05). Over-expression of TERF2 was observed in TEL-AML1 subgroup of which p-value=0.0002, (p<0.05). In contrast, under-expression of TERF2 was found in those having BCR-ABL1 fusion transcripts of which p-value=0.0221, (p<0.05). TERF2 also have found to have a better survival advantages for cALL patients. Over-expression of TERF2 is associated with good prognosis in cALL whilst under-expression is associated with poor prognosis in cALL patients.  Measurement of TERF2 gene expression allows proper stratification of cALL subtypes into its respective prognostic indicator classification.
4th International Conference on Molecular Biology and Biotechnology 2021 (ICMBB2021), 1st-3rd June 2021
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Volume 29 (3); 2021 

Analysis of DNA methylation level and mRNA expression of Transient Receptor Ankyrin Member 1 (TRPA1) in endometriosis-associated pain
Ocktariyana, Nurul Hikmawati, Andon Hestiantoro, Raden Muharam, Muhammad Luky Marwali, Agus Surur, Tiara Aninditha, Gita Pratama, Anisah Zahrah, Nabilla Farah Naura, Asmarinah
APJMBB 29 (3): 1-10
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.3.01
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Transient Receptor Ankyrin Member 1 (TRPA1) is an ion channel family protein that regulates pain sensation through
sensory neurons' activity. This study's purpose to analyzes the DNA methylation and mRNA expression level of the TRPA1 gene in endometriosis and its correlation with pain level. Twenty samples of peritoneal endometriosis and endometrial samples were obtained from women with endometriosis, which was subsequently compared to 20 endometrial samples of women without endometriosis. The DNA methylation level of TRPA1 was analyzed using Methylation-specific PCR (MS-PCR) and ImageJ software, while the mRNA expression of TRPA1 was analyzed using qRT-PCR. Furthermore, the pain level was measured using the numeric rating scale (NRS) by interviewing all the women. This study showed that there was a significant difference in the mRNA expression of TRPA1 in peritoneal endometriosis. The TRPA1 was unmethylated in both peritoneal and endometrial samples in endometriosis. However, DNA Methylation level of TRPA1 in peritoneal and endometrial of endometriosis compared to normal endometrial were no significant difference. Additionally, there was no correlation between DNA methylation level and mRNA expression level of TRPA1 in all samples, along with the endometriosis-associated pain.
Ochratoxin contamination in pig feed from pig farming centres in Indonesia
Variena Intansari, Agustin Indrawati, Sri Murtini
APJMBB 29 (3): 11-17
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.3.02
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Ochratoxin A (OTA) is the most common toxin found in nature. Ochratoxin is a metabolic product by Aspergillus spp. and Penicillium spp. OTA produced by many Aspergillus or Penicicillium species that contaminate animal feeds can cause diseases. This study aims to determine the incidence of ochratoxin contamination in pig feed such as pellets, meat bone meal (MBM), and tofu by products. The sampling method used the disease detection formula to collect 36 samples 36 taken from several regionsNorth Sumatra, East Nusa Tenggara, Bali, Bulan Island, Solo, Lampung, Banten, Bogor and Jakarta. Ochratoxin contamination in animal feed was detected based on the Enzyme Linked Immunosorbent Assay (ELISA) using Agraquant® ochratoxin assay following the manufacturer’s protocol. The analysis showed that 6 of the 36 samples (16.7%) were tested positive for ochratoxin contamination. Pellets and MBM were the feed and feedstuff which were found to contain ochratoxin. The concentration of ochratoxin in MBM was below the limit set by the National Agency of Drug and Food Control of the Republic of Indonesia (5 ppb). The high contamination of ochratoxin was found in pelletfrom Lampung (19 ppb). The fungi found to dominates the culture media and come from pellet feed were Aspergillus flavus and Aspergillus niger
Development of cinnamaldehyde loaded-alginate based film for food packaging
Siti Zubaidah Abdullah, Woei Yenn Tong, Chean Ring Leong, Syarifah Ab Rashid, Nur Amiera Syuhada Rozman, Nur Humaira Mohammad Hamid, Sabrina Karim, Najua Delaila Tumin, Siti Afifah Muda, Lily Suhaila Yacob
APJMBB 29 (3): 18-25
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.3.03
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The increased incidence of food spoilage has exacerbated the need to enhance the protection of processed foods. Food packaging incorporated with bioactive components can be exploited to improve the security of food products from microbiological contamination. Thus, this study aimed to generate an alginate-based film loaded with cinnamaldehyde for use in food packaging. Aspergillus niger ATCC 9029 was utilised to assess the biodegradability of the film. The fungal growth on the plate with alginate film was evident after 21 days of incubation period and 29.7% of weight loss was monitored. The mechanical characterisation of the film indicated that the resulting film was sturdy and flexible. In the cinnamaldehyde release test, no burst release was observed. The release was slow and gradual. Out of 8 test microorganisms, only 6 microorganisms were inhibited by the film with cinnamaldehyde during the cross streak test. The growth inhibition test signified that there were 4 Gram-positive bacteria with 100% growth inhibition. The application of the film showed a notable reduction of bacterial load in the cooked rice. Besides, it exhibited 5.0-log suppression of bacterial growth, which relative to control. The results indicated that the alginate film incorporated with cinnamaldehyde could be potentially used as an eco-friendly food packaging material.
Assessment of total bacterial diversity in whiteleg shrimps and its aquaculture environment in Pangkajene and Banyuwangi, Indonesia
Yuni Puji Hastuti, Yuli Siti Fatma, Hardi Pitoyo, Wildan Nurussalam, Jajang Ruhyana
APJMBB 29 (3): 26-37
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.3.04
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Detection of bacterial diversity in whiteleg shrimps and its rearing water is a vital first step in monitoring aquaculture activities. Bacterial community imbalance in whiteleg shrimps and its rearing water influences the quality and quantity of shrimp production. Identifying the bacterial community provides basic information related to dominant bacterial groups in whiteleg shrimps and environments, providing recommendations for proper environmental monitoring and management. In this study, we investigated bacterial community structure in the rearing water and intestinal tract of whiteleg shrimp (Litopenaeus vannamei) collected from two sites, i.e., Pangkajene, South Sulawesi (SU) and Banyuwangi, East Java (BW), Indonesia. The bacterial community was analyzed using amplicon sequencing with Illumina sequencing platform based on the V3-V4 region of the 16S rRNA genes. Bacterial diversity and composition were found differed between the rearing water and the shrimps’ intestines. Bacterial diversity in the rearing water of Banyuwangi (W.BW) was higher than that of Pangkajene (W.SU). Proteobacteria, Bacteroidetes, and Firmicutes were found as the most dominant phyla in rearing water from both farms, while distinct bacterial composition was observed in the shrimps’ intestines. The shrimp intestine from Banyuwangi (U.BW) was dominated by Firmicutes (22.36%), Proteobacteria (22.33%), and Verrucomicrobia (21.11%). In contrast, the shrimp intestine from Pangkajene (U.SU) was highly dominated by Tenericutes (88.54%), followed by Proteobacteria (4.66%), and Firmicutes (2.27%). The difference in bacterial community structure between the rearing water and shrimps’ intestines suggested that the host intestinal environment might have greater selective pressure for bacterial composition inhabiting L.vannamei intestines. Our observations suggest that the shrimps cultured in the rearing water with the similar dominant bacterial group have specific intestinal bacterial diversity.
Short communication: Canarium odontophyllum Miq. (dabai) stem bark arrested HCT 116 cell line at G0/G1 checkpoint
Dayang Fredalina Basri, Ngieng Sheng Ee, Nor Fadilah Rajab, Dharrshine Yoganathan, Raha Ahmad Raus
APJMBB 29 (3): 38-45
Article DOI: https://doi.org/10.35118/apjmbb.2021.029.3.05
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Canarium odontophyllum Miq. is an exotic plant which is native in Borneo and belong to the Burseraceae family. It contains phytochemicals such as saponin, terpenoid, flavonoid and phenolic compound with potential anticancer property. It has been found that the extract of this plant negatively affected colorectal cancer cells by stimulating apoptosis. To elucidate the apoptosis mechanism induced by the plant extract, this study evaluated the effect of C. odontophyllum stem bark acetone extract on cell cycle distribution of HCT 116 cell line using propidium iodide assay. For this purpose, IC50 of the acetone extract of C. odontophyllum was first determined by treating HCT 116 cells with the extract for 24, 48 and 72 hours. It was found that the acetone extract of C. odontophyllum inhibited proliferation of HCT 116 at IC50 value of 55.09 ± 18.29 µg/mL for 24 hours treatment, 37.81 ± 5.09 µg/mL for 48 hours treatment, and 114.9 ± 16.08 µg/mL following 72 hours treatment. Using IC50 value of 48 hours treatment, it was observed that C. odontophyllum acetone extract arrested the HCT 116 cells at G0/G1 checkpoint. Based on this result, it can be concluded that one of the apoptosis mechanisms induced by C. odontophyllum is by arresting cell cycle of HCT 116 cells at G0/G1 checkpoint. This finding warrants further investigation on how C. odontophyllum causes the cell cycle arrest and its potential to become anticancer agent.

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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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