Current Issue

 Volume 28 (1); 2020 

Nuclease pre-treatment increases efficiency of whole genome sequencing of Influenza B virus in respiratory specimens
Wudtichai Manasatienkij, Piyawan Chinnawirotpisan, Weerayuth Kittichotirat, Sriluck Simasathien, Louis R. Macareo, Damon W. Ellison, Supapon Cheevadhanarak, Wiriya Rutvisuttinunt, Stefan Fernandez, Chonticha Klungthong
APJMBB 28 (1): 1-13
Article DOI:
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The use of next generation sequencing (NGS) directly on respiratory specimens to obtain viral whole genome sequence (WGS) enhances the capability for rapid and unbiased viral characterization. One of the challenges of using NGS directly in influenza-like illness (ILI) respiratory specimens is the higher proportion of host and bacterial genome compared to viral genetic materials found, which reduces the likelihood of obtaining complete viral genome sequences. This study aims to evaluate nuclease pretreatments prior to sequencing of influenza B virus directly from ILI respiratory specimens.  Sequence data were mapped to human, bacteria and influenza B viral genome. In the absence of any nuclease pretreatments, the sequence reads identified as Haemophilus influenzae, Haemophilus parainfluenzae, Neisseria meningitidis and Veillonella parvula were the most prominent genetic materials in respiratory specimens. Filtration followed by nuclease treatment reduced bacterial sequence reads by at least 70 folds in all 4 tested samples, supporting the direct application of NGS in ILI respiratory specimens. Although the pretreatment methods significantly reduced human genome sequences, the remaining human genome especially human rRNA still impact the number and proportion of the viral sequence reads.
Potential use of Pennisetum purpureum for phytoremediation and bioenergy production: a mini review
Nurul Atiqah Osman, Ahmad Muhaimin Roslan, Mohamad Faizal Ibrahim, Mohd Ali Hassan
APJMBB 28 (1): 14-26
Article DOI:
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Organic and/or heavy metal pollutants in soil and wastewater can be remediated by phytoremediation. Phytoremediation combines the disciplines of plant physiology, soil microbiology and soil chemistry. There are several ways by which plants extract, stabilize, filtrate, volatilize or degrade the contaminants. However, the effectiveness of phytoremediation relies upon the type of plant used. Pennisetum purpureum, commonly referred to as Napier grass, is one of the exceptional phytoremediators due to its rapid growth rate and ability to survive in highly contaminated soils. In the present review, the potential use and applicability of P. purpureum to remediate various contaminated areas was highlighted and comprehensively discussed, especially the five phytoremediation mechanisms involved (i.e., phytodegradation, phytoextraction, phytofiltration, phytostabilization, phytovolatilization). The application and management of P. purpureum
in soil and wastewater phytoremediation were also critically presented. The coupling of phytoremediation and bioenergy is the zero-waste concept that can be applied since P. purpureum contains high lignocellulosic content that can be utilized as carbon source for biofuel production, such as ethanol and butanol.
Plasma miR-126 levels and its genomic polymorphism SNP rs4636297 in Type 2 diabetes
Monireh Rezai, Shirin Shahbazi, Nasrin Mansournia
APJMBB 28 (1): 27-33
Article DOI:
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Since the variation of microRNAs expression has been described in type2 diabetes, we aimed to evaluate the plasma levels of miR-126 with known roles in the angiogenesis. Genomic polymorphism of miR-126 (SNP rs4636297) was also assessed as it is involved in the processing of pri-miR-126 to mature microRNA.Genotyping of the SNP rs4636297 was carried out by PCR-RFLP using HaeII enzyme. Real-time PCR assay was applied to assess miR-126 expression, which the fold change in gene expression was calculated by the Relative Expression Software Tool (REST). Data analysis was performed using exact-like logistic regression, Fisher exact test and the elrm package with the R software. Our data indicated that the group with BMI greater than 30 was at greater risk of diabetes (odds ratio=14.4, p value=0.00), compared with the BMI=19-25. The mean fold decrease in miR-126 gene expression in diabetic samples relative to normal samples was 0.653 (95% confidence interval: 0.012-18.765). All patients with the lowest levels of miR-126, had experienced diabetic complications. Considering the AA genotype as the reference, the odds ratio of diabetic complications for the GG and AG genotype were 1.2 and 1.43, respectively. miR-126 is one of the key factors in predisposition to diabetes and diabetic complication. Since the lowest expression of miR-126 leads to the diabetic complications it can be concluded that genetic predisposing factors should be considered in the management and treatment of Type 2 diabetes.
In-silico molecular docking study of novel derivatives of erlotinib in glioma
Deepika Tripathi, Sabiha Imran
APJMBB 28 (1): 33-38
Article DOI:
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Gliomas are primary brain tumours arises from the glial cells. Gliomas associated risk factor depend upon its degree of stage and grade. Erlotinib is a quinazoline derivative and antineoplastic agent that capacities as a Protein Kinase Inhibitor for Epidermal growth factor receptor (EGFR) related tyrosine kinase and widely used for treating non-small cell carcinoma. Erlotinib, (a tyrosine kinase inhibitor) has been found in the glioblastoma. ErbB (EGF family receptor tyrosine kinase) established to control cell survival, proliferation, migration, and differentiation. The ErbB receptor accounts for the restricted and redundant functions in the maintenance of tissues in adult mammals. Newly discovered drug targets and personalised treatments could be used to identify the characteristics of any individual’s tumorigenesis. To see the binding mode of deferent classes of erlotinib carefully chosen based on the structural comparison, we employed simulated screening and molecular docking based on the theory of evolution. The study consists of a simulated screening of around 3200 molecular derivatives of erlotinib and their molecular docking using a Lamarckian genetic algorithm. Molecular docking results showed binding energy ranging from -12.15 kcal/mol to -1.17 kcal/mol. Top 10 best-docked proteins were analyzed using UCSF Chimera and finding discovered the complicated atomic-scale properties between ligand and the target protein. Further wet lab study requires to study the actual binding as binding mode provided. To identify new inhibitors of EGFR with higher potency and specificity, additional information is needed for future designs molecules.
Transcriptome analysis of Phytophthora infestans and Colletotrichum coccodes in tomato to reveal resistance mechanisms
Abbas Saidi, Zahra Hajibarat, Zohreh Hajibarat
APJMBB 28 (1): 39-51
Article DOI:
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Tomato is considered as an important commercial crop and protective food and it’s yield is extremely affected by different pathogens. Such harmful pathogens severely affecting tomato include Phytophthora infestans (oomycete) and Colletotrichum coccodes (fungi). In this study, gene expression analysis was performed using the microarray datasets available in the public database to investigate differentially expressed genes DEGs under developmental and anatomy conditions and consequently constructing the predicted PPI (Protein-protein interaction) network. Here, a comparison of abundances revealed that 51 genes were differentially expressed due to oomycete and fungi stresses in tomato. Expression analysis revealed that Solyc06g007510.3 was up-regulated whereas Solyc05g008850.3 was down-regulated in all developmental stages and anatomy conditions to manage auxin metabolism under fungi and oomycete infections in tomato. The gene ontology included cellular metabolic and organic substance metabolic processes, cellular biosynthetic, organic substance biosynthetic, and cellular macromolecule metabolic process, chloroplast, intracellular membrane-bounded organelle, heterocyclic compound binding, organic cyclic compound binding, ion binding, binding, and catalytic activity.  Analysis of cis-elements illustrated that 12 most general cis-regulatory elements (CATT-box, TATA-box, MYB, MYC, ABRE, G-box, Box 4, AAGAA-motif, MBS, as-1, TGACG-motif, and CGTCA-motif) were identified in promoter region. The highest frequency of cis-elements included CATT-box and TATA-box in response to the pathogens at different growth stages in tomato. Our findings can provide a better insight to the regulatory mechanisms involved in response to fungi and oomycete infection in tomato which can aid to improve the efficiency of developing tomato varieties.
Development of attenuated Vibrio parahaemolyticus mutant strains as potential live vaccines
Vu Thi Bich Huyen, Nguyen Xuan Viet, Pham Thi Tam, Man Hong Phuoc, Huynh Viet Tung, Nguyen Dang Quang, Do Thanh Van
APJMBB 28 (1): 52-67
Article DOI:
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Vibrio parahaemolyticus is a pathogenic Gram-negative bacterium that commonly infects numerous species of marine fishes. In this study, we obtained eight attenuated mutant strains by the selection of rifampicin-resistant V. parahaemolyticus from three wild-type strains. Among eight selected strains, four strains (L4650, A650, L4200 and A400) were widely attenuated by virulence analysis. Sequences of two genes, tlh and toxR, that related to virulence were analyzed. The results showed that seven attenuated strains were different in sequences of the tlh gene while there was no difference in sequence of toxR gene in comparison with that of wild-type strains. Three attenuated strains (L4650, L4200 and A650) had the nucleotide changes in tlh gene that resulted to the changes in the size of deduced amino acid sequences. In addition, rpoB gene coding for b-subunit of RNA polymerase that related to Rifampicin resistance was analyzed for all selected strains. Two strains (A650 and L4650) had the nucleotide changes in ropB sequence. To assess the potential use of the L4650 strain as a live attenuated vaccine, the groupers (Epinephelus coioides) were intraperitoneally injected with the L4650 strain and later challenged with the virulent wild-type strain at three different doses. Results showed the protection for L4650 strain vaccinated-fish produced relatively earlier than usual at 15 days post-vaccination with relative percent survival (RPS) values from 96.91-100%. At 60 days post-vaccination, the RPS values of vaccinated fish were 96.27-100%. Our results suggest the L4650 strain could become a potential vaccine candidate against V. parahaemolyticus for future studies.
Statistical optimization of an acid protease production by a local Aspergillus nigerMH109542 using a medium based on decommissioned dates
Malika Benkahoul, Amina Bramki, Aicha Belmessikh, Aicha Mechakra-Maza
APJMBB 28 (1): 68-76
Article DOI:
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The production of an acid protease by liquid fermentation is performed on a medium based on decommissioned dates by a local mold isolated from an extreme environment. The used mold is isolated from the thermal soil in Hammam Safsaf of Teleghma (Algeria). Phenotypic and molecular identification has shown that it is Aspergillus niger. This fungi strain exhibited exo-protease activity on milk agar. The fermentation medium based on decommissioned dates is enriched by other factors according to a statistical method; the plan of Plackett and Burman. This method (N = 8 experiments and N-1 factors) allowed the optimization of the enzyme production and the growth of the mold. The statistical analysis of the obtained results shows an increased enzyme activity (650.20U), in the presence of yeast extract and salts with probabilities p = 0.239 for the first and p = 0.190 for the second. Furthermore, the corn-steep-liquor (p = 0.229) and yeast extract (p = 0.053) present a significant effect on mold growth. In conclusion, the culture of A. niger on optimized medium gives good yields of biomass and proteolytic activity compared to the basal medium. These results are encouraging. In fact, the use of a cheap and available substrate such as decommissioned dates saves the cost price of fermentation on an industrial scale.
Mini review on phenol biodegradation in Antarctica using native microorganisms
Kavilasni Subramaniam, Siti Aqlima Ahmad, Noor Azmi Shaharuddin
APJMBB 28 (1): 77-89
Article DOI:
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Though Antarctica has once been considered as the most pristine land on earth, however, recently many literatures concluded that it is not a zone free from anthropogenic pollutants, which have been mostly associated with long-range atmospheric transport and deposition in the area. Numerous organic pollutants including phenol have been classified as the priority pollutants by the United States Environmental Protection Agency (US EPA) due to their high toxicity. The increased level of phenol concentration in the Antarctic environment poses a significant risk to the aquatic as well as terrestrial lives and public health due to its persistence, biomagnification and accumulation in the food chain. Therefore, bioremediation actions are significant to overcome this problem. Phenol degradation at cold climate needs the use of microorganisms that has the ability to thrive and function at low temperatures as well as withstand the toxicity of phenol. The utilisation of native microbes as phenol-degraders has proven the effectiveness of bioremediation even though phenol has anti-microbial properties. This paper discusses the sources and toxicity of phenol, existence and effect of phenol on the Antarctic environment, the potential method for eliminating phenol from the environment and suggestion for future prospect.
Trihoney ameliorates hypercholesterolemia-induced epididymal histopathological changes in male rabbits
Zenab B. Hamad Mohamed, Muhammad Bin Ibrahim, Hamad Abdulsalam Hamad Alfarisi, Azantee Yazmie Abdul Wahab, Azliana binti Abd Fuaat, Che Anuar Che Mohamad
APJMBB 28 (1): 690-100
Article DOI:
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Hypercholesterolemia has been linked to weight change and histopathological alteration of male reproductive organs. The epididymis was suggested to be an early target of lipid-related infertility and can be dramatically affected by excess intake of a high cholesterol diet. On the other hand, the interest has been increased towards the use of honey as a prophylactic and therapeutic agent for various diseases. Therefore, the purpose of this study is to investigate the effects of Trihoney (a mixture of Trigona, Mellifera and Tualang) on epididymal weight change and histopathological alterations in hypercholesterolemic male rabbits and compare its effects with atorvastatin. Forty-eight mature male New Zealand white rabbits were divided into 6 groups. Two groups received standard rabbit pellet with 0 and 0.6 g/kg/day of Trihoney respectively while the other four groups received 1% cholesterol diet with 0, 0.3, 0.6 g/kg/day of Trihoney, and 2 mg/kg/day of atorvastatin. After 12 weeks, the rabbits were sacrificed and the epididymides were harvested for evaluation of weight and histopathological changes. Administration of 1% cholesterol diet either alone or in combination with atorvastatin caused a significant reduction in the epididymal weight and epididymal atrophy. Supplementation of Trihoney particularly at the dose of 0.6 g/kg/day improved epididymal weight, regained the normal architecture of the epididymal histology and increased the number of mature sperm inside the tubules of the epididymis. Based on these results, Trihoney exhibited its potential health benefit as a protective agent against epididymal weight reduction and histopathological alterations in hypercholesterolemic rabbits.
Prediction of B-cell epitope by in silico analysis of Mycobacterium tuberculosis Ag85B antigen
Nihayatul Karimah, Sabar Pambudi
APJMBB 28 (1): 101-109
Article DOI:
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Mycobacterium tuberculosis (Mtb) is a causative pathogen of tuberculosis (TB) that emerges as one of the deadliest communicable diseases in Indonesia. The quest for protein biomarkers for TB has been conducted in order to develop a TB diagnostic kit and a TB vaccine. One of the abundant biomarkers in the TB infected human serum is the Ag85B antigen. In this study, we employed immunoinformatic prediction tools such as Ellipro and VaxiJen to predict the B-cell epitopes of Ag85B wildtype and multidrug resistance type (mutant). We then performed molecular docking simulation to evaluate the predicted epitopes using HADDOCK. The screening of both continuous and discontinuous B-cell epitopes using criteria-based analysis resulted in the eight linear epitopes and two conformational epitopes in Ag85B with high antigenicity. The
in silico analysis showed no major differences between Ag85B wildtype and Ag85B mutant, implying Ag85B a good target for TB vaccine candidates but not for a specific biomarker that differentiates wild-type and mutant TB.
Stevia: limiting cholesterol synthesis in Hep-G2 cells
Amirul Nazhan Ilias, Hazilawati Hamzah, Intan Safinar Ismail, Mokrish Ajat
APJMBB 28 (1): 110-119
Article DOI:
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As of today, no literature has been reported on the efficacy of stevia on lipid regulations conducted in vitro. Thus, the current study was focusing on the potential of Stevia rebaudiana bertoni as an anti-hypercholesterolemia substitute in limiting the de novo cholesterol synthesis in Hep-G2 cell line. The cytotoxicity and lipid internalization effects of stevia on Hep-G2 cells were assessed quantitatively and qualitatively in this study. As evaluated by MTT assay, commercialized stevia (0.5-20.0 mg/ml) and stevioside (1.0-10 µM) inhibited Hep-G2 cells viability in a dose-dependent manner for 24 hours. IC50 was detected at 8.68 mg/ml (commercialized stevia) and 10.91 µM (stevioside). From the assay, suitable concentrations were chosen to study the effect of stevia on cholesterol internalization in Hep-G2 cells supplemented with exogenous lipids. Cholesterol quantification assay revealed that high concentration commercialized stevia and stevioside promoted significant cholesterol internalized in Hep-G2 cells as compared to simvastatin. Finally, immunofluorescent microscopy assessment was done to qualitatively observe the formation of lipid droplets and low-density lipoprotein receptor in relation to total cholesterol extracted. The microphotographs of immunofluorescent microscopy were in parallel to results obtained from the cholesterol quantification assay which further revealed the effect of stevia as a potential anti-hypercholesterolemia agent.

Volume 28 (2); 2020

Mechanism and challenges associated with adaptation and evolution of drug-resistant bacteria: an overview
Shikha Kapil, Tarun Kumar, Vipasha Sharma
APJMBB 28 (2): 1-18
Article DOI:
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Antimicrobial resistance is one of the leading challenges in the human healthcare segment. Advances in antimicrobial resistance studies have revealed various intrinsic, adaptive or acquired factors to be involved for pathogenicity. Antimicrobialagents are either bactericidal or bacteriostatic in action and prescribed according to the mode of action. Various factors are confined for the antimicrobial activity of these agents via biochemical, mechanical, physiological and molecular mechanisms.Microbial cell expresses a number of alternates responsible for the evolution of resistance against these agent activities involving cell surface modifications, enzyme inhibitions, modifications in efflux system, protein carriers and mutations in nucleic acids. Apart from this, the successful adaptations of such microbes have also been observed with the transfer of responsible genes through miscellaneous operations such as vertical evolution, horizontal gene transfer, co-selection, compensatory and random mutation. In addition, alterations or modifications in biochemical and physiological mechanisms atcellular levels are also responsible for antibiotic resistance. This article briefly shows the present scenario of antimicrobial resistance and the alternatives to overcome this global issue in future.
Taxonomic note: speciation within the operational group Bacillus amyloliquefaciensbased on comparative phylogenies of housekeeping genes
Mohamad Syazwan Ngalimat, Suriana Sabri
APJMBB 28 (2): 19-26
Article DOI:
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Many of the publically available Bacillus16S rRNA genes and genomes in the NCBI database are inconsistently assigned as B. amyloliquefaciens. The highly conserved nature of the 16S rRNA gene makes it fail to differentiate species within the operational group B. amyloliquefaciens. Here, comparative phylogenies of the complete 16S rRNA, gyrBrpoBtrpBrecA, and cheA nucleotide sequences of bacterial strains within the operational group were analyzed. As the result, the gyrBrpoB, and trpB phylogenetic analyses showed stable topology that comprised three monophyletic clades: (i) B. amyloliquefaciens; (ii) B. siamensis; and (iii) B. velezensis. Phylogenies derived by comparison of the gyrBrpoBtrpBrecA, and cheA with the 16S rRNA gene-derived phylogeny was significant as evaluated by the likelihood ratio test. The trpB, rpoB, and trpB gene-derived phylogenies provide a tool for speciation within the operational group B. amyloliquefaciens.
Insertion/deletion of angiotensin-converting enzyme gene polymorphism as a marker of polycystic ovary syndrome (PCOS) development in South Indian cohort women
Nishu Sekar, Abilash Valsala Gopalakrishnan
APJMBB 28 (2): 27-31
Article DOI:
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Polycystic ovary syndrome (PCOS) is an endocrine/metabolic disease because of the elevated levels of androgen which could lead to anovulation. The angiotensin-converting-enzyme (ACE) is a zinc metallopeptidase that converts angiotensin I to angiotensin II. ACE is bound to the plasma membrane and expressed in many tissues such as ovarian tissues. The renin-angiotensin system (RAS help the production of angiotensin, angiotensinogen, and ACE. Angiotensin II plays a major role in ovulation, steroidogenesis, follicular atresia and hyperandrogenic syndromes such as PCOS. This study aimed to determine the association of ACE polymorphism in PCOS to analyze the distribution allele frequency of insertion or deletion variation inPCOS patients of the South Indian cohort. A total of 430 women with PCOS confirmed based on the Rotterdam criteria and 300 age and sex-matched control samples were studied. PCR technique was used to determine the frequency of polymorphism in the ACE gene. The genotyping distribution of II, DD and ID in PCOS was 4.56%, 30.23%, and 65.11%, respectively, whereas the control group showed 30%, 20%, and 50% for II, DD and ID, respectively. The deletion (D) allele frequency was 62.79% and insertion (I) allele was 37.2 % in PCOS patients, whereas in the control group, it was 45% and 55% for D and I alleles, respectively. This study concludes that the distribution of deletion (D) allele frequency of ACE could be considered as a genetic marker for PCOS in the South Indian cohort.
Microencapsulation of Bifidobacterium animalis subsp. lactis BB-12 with mannitol
Agnes Kai Lin Yong, Ka Wai Lai, Hasanah Mohamad Ghazali, Lee Sin Chang, Liew Phing Pui
APJMBB 28 (2): 32-42
Article DOI:
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Bifidobacterium animalis subsp. lactis BB-12 (BB-12) was microencapsulated using co-extrusion technology with chitosan coating and the incorporation of mannitol as prebiotic. Optimization of coating material chitosan concentration (0–0.5% w/v) and mannitol concentration (0–5% w/v) as prebiotic were performed to determine the formulation that produces beads with desired properties. The microencapsulation efficiency (MEE) of free and microencapsulated BB-12 (with and without mannitol) were determined. All forms of BB-12 further subjected to sequential digestion in simulated gastric juice (SGJ, pH 2.0) for 2 hours and simulated intestinal juice (SIJ, pH 7.5) for 3 hours. The results indicated that 0.4% (w/v) of chitosan coating and 3% (w/v) of mannitol were the optimum concentrations to produce microencapsulated BB-12 with the highest MEE of 89.15% and the average bead size of 805 µm. The BB-12 beads produced through co-extrusion were spherical with a smooth surface. Throughout the five hours sequential gastrointestinal digestion, both microencapsulated BB-12 with and without mannitol were able to maintain their viable cell count at least 10CFU/g at the end of the incubation. The presence of prebiotic mannitol showed a significant protective effect on the microencapsulated BB-12 during gastrointestinal transit.
Analysis of mRNA expression and DNA methylation level of RAC1 gene encoding focal adhesion molecule in endometrial and peritoneal endometriosis
Irwina Eka Deraya, Andon Hestiantoro, Raden Muharam, Muhammad Lucky Marwali, Agus Surur As'adi, Darmawi, Achmad Kemal Harzif, Gita Pratama, Ocktariyana, Annisah Zahrah, Asmarinah
APJMBB 28 (2): 43-49
Article DOI:
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Recent studies showed that the expression of RAC1, a gene encoded focal adhesion molecule, was predominantly found in endometriosis. As gene expression may be regulated by DNA methylation. Therefore, this study aimed to analyze promoter methylation level of RAC1 gene and mRNA expression in endometrial and peritoneal endometriosis tissues. This study using 20 endometrial and 9 peritoneal tissues from the same patients and 20 normal endometrial. The DNA and RNA from samples were isolated, DNA was converted using sodium bisulfite and amplified using Methyl Specific Polymerase Chain Reaction (MSP) method. The methylation level was determined by the intensity measurement of the bands that arose in gel electrophoresis using ImageJ software, whereas mRNA expression level was measured by Reverse Transcription-quantitative PCR (RT-qPCR) method. The mRNA expression level of RAC1 gene in peritoneal endometriosis increased compared to normal endometrium, as well as compared to endometrial endometriosis, but there was no significant difference in endometrial endometriosis compared to normal. Promoter hypermethylation level of RAC1 gene in peritoneal endometriosis was significantly different compared to normal endometrium, however not significant to endometrial endometriosis. Methylation level of its gene in endometrial endometriosis shown no significant difference compared to normal. There was association between promoter hypermethylation level and its mRNA expression in endometrial endometriosis (R= 0.014; p=0.952). The elevation of mRNA expression of RAC1 gene plays a role in endometrial cell migration to peritoneum, and associated with promoter hypermethylation level of its gene.
Effect of Acacia purified tannins extract and polyethylene glycol treatment on in vitroruminal fermentation pattern and methane production
Khaoula Khelalfa, Rabah Arhab, Antonio Ignacio Martín-García, Nabila Zaabat, Alejandro Belanche
APJMBB 28 (2): 45-62
Article DOI:
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Condensed tannins are polyphenolic compounds which can exert beneficial effects in ruminants. They have the ability to bind proteins and decrease their degradation. They have also been reported to reduce methanogenesis and improve ruminants performances. The present work aimed to study the effect of purified condensed tannins from Acacia horrida extract on fermentation parameters, gas and methane production. In this context, seven substrates were selected: four Acacia species (Acacia pycnantha, Acacia dealbata, Acacia horrida and Acacia cyanophylla) and three plant by-products (date palm leaves, grenade peel and artichoke stems). Biological activity of tannins was evaluated by the incubation of the substrates in vitro with Polyethylene glycol (PEG). Tannins decreased concentration of all fermentation parameters (gas (p < 0.0001), methane (CH4) (p= 0.0983), ammonia (N-NH3) (p= 0.0382), Volatile Fatty Acids (VFA) (p= 0.0009), acetate (p <0.0001), propionate (p= 0.1024), butyrate (p= 0.0373), isobutyrate (p= 0.0046), valerate (p < 0.0001), isovalerate (p= 0.0032)). Substrates were also characterized by their content in main nutrients (dry matter (DM), organic matter (OM), ash, crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) ,acid detergent lignin (ADL)) and in secondary metabolites (total phenols (TP), total tannins (TT) and condensed tannins (CT) ). All substrates presented high CP content except by-products (> 100 g/kg DM) and moderate cell-wall components. CT content was comprised between 120.5 and 680.4 g/kg DM in plants and between 23.1 and 170.4 g/kg DM in plant by-products. A. horrida presented the highest biological activity. Thus, their CT were extracted and purified on a sephadex LH-20 column. Purified tannins from A. horrida were incubated with two different forages alfalfa hay and barley straw at three different concentrations: 50, 100, 150 mg/g DM. Results showed that purified tannins from A. horrida had no effect on fermentation parameters (P > 0.05). The effects of A. horrida tannins may be strongly linked to their structure and their molecular weight more than to their concentration. For this reason, it is interesting that this work may be completed by physical characterization of these tannins.
Assessment of growth and phytochemical quality of Gynura procumbens through nitrogen, potassium fertilization and evapotranspiration replacement interaction
Mohamad Fhaizal Mohamad Bukhori, Hawa ZE Jaafar, Ali Ghasemzadeh, Uma Rani Sinniah, Gayatri Karipaya, Keezrawati Mujan Yusuf
APJMBB 28 (2): 63-91
Article DOI:
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Despite the progressive reports on potential pharmacological properties of G. procumbens, the importance of agronomic requirements to produce high yields and phytochemical content that may vary due to environmental variations are often overlooked. Therefore, this study was conducted to examine the effects of N, K and ER interaction on the growth and phytochemical content of G. procumbens. The study was a three-factorial experiment; two rates of N and K, four rates of ER and three H time.  The treatments have affected plant growth and phytochemical content significantly (p≤0.05) with stronger effect on physiology and biochemical attributes (p≤0.01). The study has shown discrete effect on growth, physiology, and phytochemicals content with N0K30>N90K0 and ER 75>50>100>25% treatment. The highest and lowest yield of plant biomass and phytochemical were observed under N0K30(70) and N90K0(25), respectively. The results have shown that the interaction and effect of treatments are highly significant (p≤0.0001) in Cond, CNB, TChlC, TPrC and TFC analysis, (p≤0.05) in NoL, CF and PWP, and not significant in TLA, Photo, TCC, TLC and TPC. The 75% ER has produced significant output of biomass as well as phytochemical content. The study also showed that low rate of N, moderate rate of K with 75% ER have produced high biomass as well as phytochemical content. Meanwhile, caffeic acid and kaempferol were demonstrated as the lead secondary metabolite compounds in this study.
Reproductive development of the priapium fish Neosthetus lankesteri Regan, 1916 (Atheriniformes: Phallosthethidae) from Pranburi river estuary, Thailand using the histological approach
Amphornphan Palasai, Sinlapachai Senarat, Phakorn Na Lampang, Niwat Kangwanrangsan, Wannee Jiraungkoorskul, Diógenes Henrique de Sequiera-Silva, Jes Kettratad
APJMBB 28 (2): 92-104
Article DOI:
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Reproduction in the family Phallostethidae is an area that has long been of interest especially in gonadal structure and gametogenesis. However, no published data concerning as mentioned above in Neostethus lankesteri Regan, 1916 from Thailand is noted. This fish is important for rural livelihood in the estuarine fisheries of Pranburi River estuary, Thailand. The construction of the female reproductive system of N. lankesteri composed of the ovary and short oviduct. The mature ovarian morphology is a single organ, which is histologically considered to be an asynchronous development ovary. The differentiating oocytes were observed and classified into oogonia proliferation, primary growth phase, secondary growth phase, post-ovulatory phase and atretic follicle phase.  Under the ovarian differentiation, our observation indicated that the ovarian parenchyma of N. lankesteri during 1.8 – 2.4 cm in standard length (SL) was accompanied by the secondary growth phases. Identification of the post-ovulatory phase was detected during our sampling periods, indicating to entire life in the Pranburi River estuary as well as true resident species. In the male reproductive system of N. lankesteri, the mature testicular parenchyma was organized morphologically as a single organ and histologically appeared to be a restricted spermatogonial lobular testis type. The differentiation of the spermatogenetic stage was composed of three phases including spermatogonial, spermatocyte and spermiogenetic phases. It was to identify possible trends that the size at first maturity of the female fish was estimated at 1.8 cm SL.  Certainly, results from this study clearly showed on the reproductive pattern of N. lankesteri, which could be applied to further studies (the reproductive cycle, spawning season and histopathology).
Identification of genes involved in flowering in Stevia rebaudiana using expressed sequence tags (ESTs)
Amin Razali, Nurul Hidayah Samsulrizal, Zarina Zainuddin
APJMBB 28 (2): 105-112
Article DOI:
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Stevia rebaudiana, or stevia, is a perennial herb native to the northern region of South America. It is commercially important due to the high level of non-caloric sweetening compound, which has been consumed worldwide as food and medicine. Stevioside is considered as the most important compound of steviol glycosides that functions as the main sweetener. Exposure of the stevia plant to long-day conditions (increased exposure to light) will delay the flowering process, which led to an increase in glycoside accumulation. The bioinformatics analysis of 5548 expressed sequence tags (ESTs) obtained from dbEST (NCBI) using the Blast2GO software and manual curation identified 7 ESTs that contain domains involved in the flowering process. The domains were the heat shock protein (Hsp) domain, squamosa promoter-binding protein (SBP) domain, CONSTANS, CO-like, and TOC1 (CCT) domains; K homology (KH) domain; and ubiquitin-conjugating enzyme domain (UBC).
Phytochemical analysis of ionic liquid-Graviola (Annona muricata) fruit extract and its acute toxicity on zebrafish early-life stages
Djabir Daddiouaissa, Azura Amid, Syahida Ahmad, Ahmed A. M. Elnour
APJMBB 28 (2): 113-124
Article DOI:
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Annona muricata, commonly known as soursop and Graviola, is a member of the Annonaceae family. Some of its phytochemicals were reported to have a neurotoxicity effect causing neurodegenerative diseases. However, different parts of this tree have been used for ages in traditional medicine due to their biological activities, such as anti-inflammatory, antimicrobial, antioxidant and anticancer effects. This study aimed to qualitatively screen the crude ionic liquid-Graviola fruit extract (IL-GFE) phytochemical composition, assess its acute toxicity and determine the lethal concentrations using zebrafish (Danio rerio) embryos. IL-GFE contains acetogenins, alkaloids, phenols, terpenoids, tannins and flavonoids. Acute toxicity effects of IL-GFE on zebrafish embryos were observed from 24 to 120 hours of post fertilisation (hpf). The survival rate, LC50, sublethal endpoints and effect of IL-GFE on the heart rate of zebrafish larvae were assessed. Results showed that the lethal concentration (LC50) of the crude IL-GFE was 173.45 μg/mL. Interestingly, no significant changes on the morphology of the treated zebrafish were observed at a concentration of 125 μg/mL. However, the heart rate of zebrafish larvae at 96 hpf was significantly decreased by 33.76% after treated with crude IL-GFE at 125 µg/mL (119.00 ± 4.72 beats/min) as compared to the untreated group (179.67 ± 4.66 beats/min). This preliminary finding showed that crude ionic liquid-Graviola fruit extract and its phytoconstituents might have the potential to be developed as a food supplement or herbal product. However, further tests need to be conducted to evaluate its medicinal properties and adverse effects on organisms of higher orders.

Volume 28 (3); 2020

Anti-inflammatory potential of the aqueous extract of Cyamopsis tetragonoloba against the MSU-induced arthritis in female Wistar albino rats
Jerine Peter Simon, Ram Kumar Katturaja, Arunraj Namachivayan, Sangeetha Nithyanandham, Manisha Parthasarathy, Sabina Evan Prince
APJMBB 28 (3): 1-12
Article DOI:
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Objective: Gout is an acute inflammation with joint swelling and pain. Uric acid plays a major role in causing gouty arthritis. Crystal shaped uric acid was found in the joint fluid at the severe condition. Guar gum is an herbal product with numerous medical benefits and its botanical name is Cyamopsis tetragonoloba. This study aims to find the protective activity of Cyamopsis tetragonoloba against the MSU-induced gouty arthritis in female Wistar albino rats. Methods: The parameters like weight assessment, paw volume, biochemical parameters (urea, uric acid and creatinine), antioxidant activities and histopathological of paw have been analyzed. The animals have been divided into 5 groups as follows 1) normal control, 2) Monosodium Urate Crystal (MSU)-induced on 1st day, 3) MSU-induced and treated with 500 mg of Cyamopsis tetragonoloba, 4) MSU-induced and treated with 1000 mg of Cyamopsis tetragonoloba and 5) MSU-induced and treated with indomethacin. Results: Aqueous extract of Cyamopsis tetragonoloba has shown reduced paw volume, renal markers, lipid peroxidation and acid phosphatase, whereas it raised anti-oxidants levels and diminished the inflammation in rats paw bone. Conclusion: The aqueous seed extract of Cyamopsis tetragonoloba has shown good benefits against MSU-induced gouty arthritis. The molecular studies have to be studied to known this mechanism in treating the inflammation.
Chromatographic separations and characterization of bioactive secondary metabolites of Apergillus quadrilineatus (MH109538) fungal strain
Amina Bramki, Edwin de Pauw, Meriem Frahtia, Andrea McCann, Gabriel Mazzucchelli, Laid Dehimat
APJMBB 28 (3): 13-21
Article DOI:
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In the interest of searching new antibiotic substances, the antibacterial effect of Aspergillus quadrilineatus fungal strain (MH109538) has been evaluated against sensitive bacteria (S. aureus, E.  faecalis, B. subtilis and E. coli) and resistant bacteria (P. aeruginosa and K. pneumonia), following the disks technique. The obtained result confirms the capacity of this strain to produce antibacterial molecules. In order to analyze and characterize these bioactive metabolites present in the crude extract, which was fractionated by column chromatography. The fractions were then separated by thin-layer chromatography. The detected spots were sampled and tested against the most sensitive bacteria (S. aureus). The results have revealed that only one fraction showed an antibacterial activity. This active fraction was analyzed by liquid chromatography coupled with mass spectrometry (LC/MS) in order to obtain the elemental composition of the present compounds. The molecular ions fragmentation (LC/ MS/MS) were used to determine the deployed formula of these compounds. The analysis of the results by Xcalibur 2.2 software reveals the presence of 24 molecules, and this, based on the retention times of the peaks obtained in the chromatogram, where, the m/z ratios, the formulas, as well as the intensities of the various compounds detected are determined.
K-ras peptide mimotope induces a humoral immune response against G12V K-ras antigen in BALB/c mice
Winfrey Pui Yee HooPui Yan Siak, Nur Aqlili Riana Alias, Jia Jia Wong, Ee Wern Tan, Adelene Ai-Lian Song, Raha Abdul Rahim, Lionel Lian Aun In
APJMBB 28 (3): 22-35
Article DOI:
Click here to download [PDF] [Supplemental Data]
Background. KRAS mutations are highly prevalent in pancreatic, lung, and colorectal carcinomas with G12V point substitution being one of the most prevalent mutations. While developments of peptide vaccines for KRAS(+) cancers are usually associated with poor immunogenicity, coupling mutant K-ras vaccines with universal CD4+ carrier molecules may enhance its outcome. Additionally, recent immunotherapeutic advances also suggest the possibility of inducing mucosal immunity against cancers using Lactococcus lactis as a live gastrointestinal delivery vehicle. Methods. A region of wild-type K-ras peptide was previously modified with a V7D substitution flanking the G12V mutation, generating a K-ras peptide (termed 68-V) with improved predicted antigenicity. This peptide was fused with a diphtheria toxoid sequence, and cloned into pNZ8048 vector within Lactococcus lactis NZ9000. BALB/c mice were then immunized orally, and then subjected to T/B cells immunophenotyping, as well as IgG and IgA detection. Results. Modified 68-V K-ras peptide and controls were successfully cloned and detection of His-tagged proteins expressed following induction by nisin was observed. Populations of CD3-CD19+ immune cells increased following immunization of 68-V, while K-ras specific-IgG and -IgA sera titers were elevated compared to wild-type and G12V K-ras controls. Conclusion. 68-V K-ras mimotope was shown to induce humoral-mediated immunity, highlighting the ability of an additional mutation flanking the G12V KRAS mutation to induce B cell activation and production of K-ras specific antibodies, while diphtheria toxoid was unable to stimulate an enhanced response when fused to 68-V. Nevertheless, these findings showed that further assessments are required to understand the role of K-ras specific antibodies within a KRAS(+) environment.
Diabetic wound infection: A review on microbial population and infection control
Nur Amiera Syuhada Rozman, Woei Yenn Tong, Chean Ring Leong, Wen-Nee Tan, Syarifah Ab Rashid
APJMBB 28 (3): 36-43
Article DOI:
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A diabetic foot ulcer is one of the major complications of diabetes and it leads to lower extremity amputation in patients. This review explores the current research on microbial populations on diabetic wounds, and also treatment alternatives to combat the infection on chronic diabetic wounds. The microbial communities exist in diabetic wound infection are diverse. Microbes rarely survived in single species of planktonic cells. They usually exist in a complex polymicrobial biofilm population which consists of different types of microorganisms. Furthermore, the development of bacterial biofilm on the wound that usually consists of multidrug-resistant pathogens also delayed the wound healing. To overcome this problem, many types of modern wound dressing were developed including hydrocolloid, hydrogel, alginate and collagen wound dressing. Besides, modern biotechnological advancements such as cell therapy, bioengineered skin, dermal scaffolds, tissue-engineered artificial skin and growth factors were also employed to promote the recovery of the wound. In conclusion, diabetes mellitus is a major health care challenge worldwide. Diabetic patients are at risk for developing foot ulcer which ultimately leads to amputation; hence a safe and effective alternative treatment is required to improve diabetic patients’ quality life.
Enhancement of very high gravity bioethanol production via fed-batch fermentation using sago hampas as a substrate
Nur Adila Muradi, Dayang Salwani Awang Adeni, Nurashikin Suhaili
APJMBB 28 (3): 44-51
Article DOI:
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Very high gravity (VHG) ethanolic fermentation is a promising technology used for producing bioethanol. However, the technology is often associated with the excessive amount of glucose that is entirely supplied in the beginning of the culture causing the fermentation process to be sluggish and therefore inhibits complete utilisation of glucose. The high concentration of glucose in the fermentation medium also elevates the osmotic pressure, which has a destructive effect on yeast cells. This study aims to enhance the production of VHG bioethanol from sago hampas hydrolysate (SHH) via fed-batch fermentation. The fermentations were performed in a 2-L stirred tank bioreactor. Batch fermentation was conducted as a control. Our results showed that the maximum yeast cell concentration achieved was significantly improved by 1.5-fold when the fermentation was carried out in fed-batch mode. The ethanol yield attained in the fed-batch culture represents an enhancement of 22% over that achieved in the batch culture. Moreover, the ethanol productivity achieved in the fed-batch culture was found to be increased by 1.8 times in comparison to the productivity attained in the batch culture. In general, this work provides useful insights into promising techniques for enhancing VHG fermentations in the stirred tank bioreactor employing agricultural residues as feedstocks.
Antibiotic resistance of ESKAPE bacteria in acute pharyngitis patients from private clinics in central Malaysia
Christina Shook Cheng Chong, Zhuang Mian Bo, Wei Keat Tan, Seshatharran Parmasivam, Shu Ting Pang, Benedict Lim Heng Sim, Christopher Kwong Chong Lee, Renee Lay Hong LimCrystale Siew Ying Lim
APJMBB 28 (3): 52-58
Article DOI:
Click here to download [PDF] [Supplemental Data
Worldwide, the ESKAPE group of pathogens - Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter species - are emerging as important pathogens which have acquired resistance towards different classes of antibiotics. Malaysia has also reported high levels of antibiotic resistance among these pathogens, based on prevalence data from hospital patients. However, the prevalence and antibiotic resistance rates of ESKAPE group members in the community are largely unknown. Therefore, this study focuses on acute pharyngitis (AP) patients from private primary care clinics in the Klang Valley, central Malaysia, as AP is the most common infection-associated reason for contact with private primary healthcare providers. Out of 205 patients, 119 were found to harbor ESKAPE group bacteria, where a total of 175 isolates were subjected to antibiotic susceptibility testing. Our results show that the most prevalent ESKAPE group member in this cohort is S. aureus, with one isolate found to be multidrug-resistant towards penicillin V, cefoxitin, erythromycin and azithromycin. We also report the first finding of community acquired resistance to cefotaxime in K. pneumoniae, imipenem in Enterobacter spp. and tetracycline, also in Enterobacter spp., in Malaysia, which were not derived from samples from a hospital setting. In conclusion, this study, which describes antibiotic resistance of community acquired ESKAPE pathogens in the Malaysian private primary healthcare setting highlights the need for a comprehensive centralized reporting system for community acquired antibiotic resistance to complement the Malaysian National Surveillance of Antibiotic Resistance.
Analyses of yield related agronomic traits of Malaysian rice varieties
Nur Aini Mohd Kasim, Chui-Yao Teh, Parameswari Namasivayam, Nur Fatihah Mohd. Yusoff, Chai-Ling Ho
APJMBB 28 (3): 59-74
Article DOI:
Click here to download [PDF] [Supplemental Data 1] [Supplemental Data 2]
Analyses of morphological and agronomic traits not only provide important information in the selection of rice varieties for breeding, these data are also important in deciding plant materials for molecular characterization and analysis of phenotypic traits of interest. The aim of the present study was to collect, analyze and compare the agro-morphological data of 6 upland rice and 17 lowland rice varieties (Oryza sativa L.) from the Malaysian rice germplasms grown under the same greenhouse condition. The common factors that contributed to yield per plant in both upland and lowland rice varieties are plant height and percentage of filled grains. Panicle length was positively correlated to grain yield in upland rice varieties while many other agronomic traits could affect the yield per plant in lowland rice varieties such as total number of tillers, total number of panicles per plant, flag leaf width and days-to-flowering. The upland rice varieties have a slightly higher average total grain number per panicle in compensation of a low number of panicles. The compensation in total grain numbers per panicle was not sufficiently significant to affect the total yield per plant most probably due to a low total number of grains per plant. Despite a high 100-grain weight in general, some of these upland rice varieties have more than 50% percentage of unfilled grains. Data on these rice germplasms may offer information on a valuable gene pool which can be utilized in different varietal improvement/development program in future.
Trihoney improves testicular weight change and histopathological alterations in hypercholesterolemic rabbits
Zenab B. Hamad Mohamed, Hamad Abdulsalam Hamad Alfarisi, Azantee Yazmie Abdul Wahab, Azliana binti Abd Fuaat, Che Anuar Che Mohamad, Muhammad Bin Ibrahim
APJMBB 28 (3): 75-87
Article DOI:
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Histopathological examination of testicular tissue is the most reliable and sensitive method for detecting effects on spermatogenesis. Hypercholesterolemia reduces testicular weight, induces testicular degenerative changes, impairs spermatogenesis, affects Leydig and Sertoli cells and induces inflammation and fibrosis of testicular tissue. Based on numerous studies, honey has the ability to improve testicular histopathological abnormalities. To date, whether honey has any protective role against the effects of hypercholesterolemia on male reproductive functions is yet to be explored. This study investigated the effects of Trihoney (a mixture of Trigona, Mellifera and Tualang honeys) on changes in testicular weight and histopathological alterations induced by hypercholesterolemia in male New Zealand white rabbits. These changes were compared with the effects of atorvastatin (a lipid lowering agent) based on the same parameters. Forty-eight male New Zealand white rabbits were assigned into 6 groups and received different diets as follows; Control: commercial pellet; CH: commercial pellet and 0.6 g/kg/day Trihoney; HCD: 1% cholesterol diet; DH1: 1% cholesterol diet and 0.3 g/kg/day Trihoney; DH2: 1% cholesterol diet and 0.6 g/kg/day Trihoney; DAt: 1% cholesterol diet and 2 mg/kg/day atorvastatin. After 12 weeks, blood samples were collected for lipid analysis, the rabbits were sacrificed and the testes were harvested to evaluate any weight and histopathological changes. Administration of 1% cholesterol diet either alone or in combination with atorvastatin caused a significant reduction in the testicular weight, testicular tubular degenerative changes and spermatogenesis impairment. Trihoney, particularly, at the dose of 0.6 g/kg/day improved testicular weight, ameliorated the testicular tubular degenerative changes and enhanced spermatogenesis. The findings of this study suggest that Trihoney plays a favourable role in the protection against testicular weight reduction and histopathological changes induced by hypercholesterolemia. On the other hand, atorvastatin per se may have toxic effects on testicular tissue.
The anticancer molecular mechanism of Carnosol in human cervical cancer cells: An in vitro study
Rand R. Hafidh, Ahmed S. Abdulamir
APJMBB 28 (3): 88-98
Article DOI:
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Carnosol, a phenolic diterpene, is one of the effective anticancer agents naturally occurring in rosemary, sage, parsley, and oregano. The chemoresistance problem increased with the routinely used chemotherapy. Therefore, the efforts to find a substitute with safe and low cost have become crucial worldwide. The current study attempts to inspect the anticancer molecular mechanisms of Carnosol on modulating up- and down- regulation of multiple genetic carcinogenesis pathways. The cytotoxicity of Carnosol on Hela cells was evaluated by MTS assay. Flow cytometry was used to assess apoptosis and cell cycle arrest. The apoptotic morphological changes were obvious by dual apoptosis assay. The differential gene expression after treatment with Carnosol was investigated by qRT-PCR. Up to 80% of the treated cells with Carnosol IC50 underwent apoptosis. Apoptosis together with cell cycle arrest in G0/G1 phase were induced significantly after treatment with Carnosol IC50. Fifteen out of nineteen genes studied were found to be remarkably up- or down- regulated after treatment with Carnosol. Six up-regulated genes (EREG, FOS-2, ID2, CRYAB, DUSP5, and TICAM2) and nine down-regulated genes (FN1,KRAS2, CCNB1-1, FEN1, MCM4, MCM5, GTSE1, CXCL1, and RALA) were recorded. These genes are candidates for future research for elucidating anticancer molecular targeted therapies, cancerous signaling and cancer development pathways in cervical cancer; moreover, elucidating the role of apoptosis, inflammation, cell proliferation, and cell differentiation in the development of cervical cancer.
Method for purification of collagen: A systematic review
Nursyahidatul Azwa Awang, Azura Amid, Zatul Iffah Arshad
APJMBB 28 (3): 99-112
Article DOI:
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Collagen has diverse applications in various industries and thus a various method of purifications has been developed over the years to produce high yield and high purity of collagen to fulfill collagen market demand. The objective of this qualitative systematic review is to summarize the current literature on the existing purification methods for collagen from different sources. Toward this end, three databases were searched and articles were screened for all original articles explaining the purification of collagen regardless of their main sources. After the comprehensive screening, out of 715 articles, 12 articles were chosen and were further reviewed. The analysis of the technical details, theory, advantages, and disadvantages of the techniques used are reported in this study. We found four types of purification methods that commonly used to purify various types of collagen from mammals, marine, bacteria, and recombinant collagen. The correlations between the prevalence of the method and the efficiency of collagen purification were also identified. It highlighted that many factors should be considered before choosing any method of purification such as types, sources and structure of the collagen itself. Affinity chromatography commonly used for purification of recombinant collagen as they have been genetically modified and appended by affinity tags whereas conventional purification method is preferable for non-recombinant collagen from mammalian and marine sources.
The influence of metal on the performance of 2,4,5-triphenylimidazole as an inhibitor of dengue virus replication
Sri Sumarsih, Muthia Dewi Pratiwi, Indah Nur Ainni, Hardinata Rachmad Sinatriya, Soegeng Soegijanto, Teguh Hari Sucipto, Harsasi Setyawati
APJMBB 28 (3): 113-121
Article DOI:
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Dengue is a contagious disease caused by dengue virus and transmitted to humans by the bite of infected Aedes aegypti mosquito. Imidazole and its derivatives are proven have ability impairing dengue virus. One of potential imodazole’s derivatives is 2,4,5-triphenylimidazole (TPI). The presence of metal to the 2,4,5-triphenylimidazole (TPI) structure through a complex compound formation highly contributes to their ability as an inhibtor dengue virus replication. Iron, cobalt and zinc were used as an ion center in the complex compound. Complex Zn-TPI and Fe-TPI showed low cytotoxic effect at all the evaluated concentrations (viability > 50%). Complex Co-TPI showed reduction of DENV-3 growth, at the lowest concentration (6.25 µg/ml) exhibited the antiviral activity (DENV-3 reduction 43%). For Fe-TPI and Zn-TPI, the reduction values of DENV-3 were 56% and 54.9% respectively.

 Volume 28 (4); 2020 

Anthocyanin-loaded polymeric dextran nanoparticle as an anti-proliferative agent on human liver carcinoma cells
Siti Zubaidah Abdullah, Chean Ring Leong, Woei Yenn Tong, Jyh Chyang Pang, Wen-Nee Tan, Syarifah Ab Rashid
APJMBB 28 (4): 1-10
Article DOI:
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In this communication, anthocyanin-loaded dextran nanoparticles from Clitoria ternatea was synthesized and characterized to test its anti-proliferative activity on the human HepG2 liver cancer cell lines. By using dextran as an encapsulant polymer, the nanoparticles appeared to be spherical, with an average size of 45.5 ± 11 nm. The surface charge of the anthocyanin-loaded dextran nanoparticle was -4.39 mV, which slightly relative to free anthocyanin (-4.46 mV), which indicate good dispersion stabilities. The Fourier transform infrared analysis showed that the anthocyanins from C. ternatea
was successfully encapsulated in dextran nanoparticles. Overall, the percentage of drug encapsulation efficiency was 3.03%. Based on the stability test, the anthocyanin-loaded dextran nanoparticle showed significantly better color stability index compared to free anthocyanin, particularly at the presence of light and temperature of 37°C and 50°C. In the anti-proliferation assay on HepG2 liver cancer cell lines, the viability of the cancer cells was significantly reduced after treatment with the anthocyanin-loaded dextran nanoparticle. The anti-proliferation activities of the nanoparticles were significantly better than free anthocyanin. Our findings revealed the ability of the anthocyanin-loaded dextran nanoparticle, in particular from C. ternatea, as an effective anti-proliferative agent against cancer cells. Nanoencapsulation with dextran significantly improve the efficacy and stability of the anthocyanins. Further investigations should be done to evaluate the
in vivo efficacy.
Effects of double emulsification on Lactobacillus plantarum NBRC 3070 stability and physicochemical properties of soursop juice during storage
Safiah Sabrina Hassan, Intan Nabihah Ahmad Fadzil, Hifa Nazirah Mohammed Yazid, Anida Yusoff, Khalilah Abdul Khalil
APJMBB 28 (4): 11-25
Article DOI:
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Sufficient number of probiotics in product are required to confer its health benefits to consumers. However, the viability of probiotics can detriment during processing. An approach of emulsion method to provide a physical barrier of probiotics against adverse environmental conditions has received considerable level of interest. Various matrices were used to emulsified probiotics in food product. However, the usage of water/oil/water (W1/O/W2) base to emulsify probiotics for beverages application remained scarce. Thus, this study was to determine the suitability of using W1/O/Wbase to emulsify Lactobacillus plantarumNBRC 3070 to be incorporated into pasteurized soursop juice (Annona muricata L.). Hence, emulsion efficiency (EE%) of L. plantarum NBRC 3070 in W1/O/W2 base, the stability of emulsified cells (log10 CFU/mL) and physicochemical changes of soursop juice (pH, titratable acidity, total soluble solid, viscosity, colour, sensory) during storage were determined. In this study, emulsified L. plantarum NBRC 3070 (109 CFU/mL) was incorporated into soursop juice and stored at 4°C for four weeks. Results obtained with 86.02% ± 0.69 emulsion efficiency (%) of L. plantarum NBRC 3070. Insignificant changes (p>0.05) were notified from viability of emulsified probiotics, pH, titratable acidity and viscosity during storage. Meanwhile, satisfactory parameters (colour, sensory and coliform counts) were obtained at the satisfactory level. As for the conclusion, W1/O/W2 base was able to maintain the survivability of L. plantarum NBRC 3070 in soursop juice during storage and quality of the product. Optimization of parameters such as water/oil compositions and storage time can be further explored to enhance product quality.
Expression and characterization of family 40 Carbohydrate Binding Module (CBM) from Vibrio cholerae Non-O1 sialidase
Gogula Selvi Asang, Shadariah Mamat, Nadiawati Alias, Asmad Kari
APJMBB 28 (4): 26-38
Article DOI:
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Carbohydrate Binding Module (CBM) is a non-catalytic protein domain found in carbohydrate-active enzyme (glycoside hydrolase) and its role is to bring carbohydrates in close proximity to the enzyme catalytic site for complete hydrolysis. The removal of this CBM from most protein domains often leads to reduced enzyme activity and efficiency. In this study, a gene encoding for family 40 CBM from Vibrio cholerae Non-O1 sialidase was cloned and successfully expressed in E. coli BL21 (DE3) strain. The CBM40 encoded 195 amino acids with 585 bp of nucleotide sequence. The protein was successfully expressed at 18°C when induced with 1 mM IPTG. Maximum expression was achieved at 20 hours after post-induction time. For purification of the protein, an anionic denaturing detergent method was used containing 1% SDS and 0.1% sarkosyl with gradient affinity elution at 50 mM imidazole concentrations. SDS-PAGE analysis of the purified CBM40 protein displayed a protein band with a molecular mass of 21 kDa. Protein characterization showed optimum stability in 100 mM citrate buffer pH 5.5, with the highest Tm value of 40 °C. The protein was stable between pH 5.5–6.2 and able to retain its activity at 27–56°C. The addition of Mn2+ and Mg2+ increased the protein melting temperature to 56°C. Meanwhile, the addition of reagents, such as 1% SDS and 1 M urea increased the protein melting temperature (Tm) to approximately 55°C. Protein stability can be influenced by many factors, including different buffers, pHs, temperatures, ionic strengths, and chemical reagents used in a study. The optimum characterization conditions established would further lead to the discovery of CBM40 protein true potential in enhancing substrate binding affinity and protein-carbohydrate recognition, which underpins its broad applications in biotechnology and protein engineering fields.
Effect of storage and frying times on stability of acrylamide and 5-hydroxymethylfurfural in fresh and frozen curry puff skins
Rasyiqah Ridzal, Norfahana Abd-Talib, Emmy Liza Anak Yaji, Kelly Yong Tau Len, Nadia Razali, Khairul Faizal Pa'ee
APJMBB 28 (4): 39-50
Article DOI:
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Acrylamide and 5-hydroxymethylfurfural (HMF) are potential compounds that are known to be neurotoxic, mutagenic, andcarcinogenic when exposed to humans. These components are formed during Maillard reaction at which sugar is reduced and specific amino acids react at high temperature, and thus producing brown colour and flavour in food. Therefore, the aimof this study was to observe the formation level of acrylamide and HMF in fresh and frozen curry puff skins. This study investigated the effects of frozen storage and frying times on the formation of acrylamide and HMF in curry puff skin. Frozen curry puff skins were stored at -18°C for 0, 7, 14, and 28 days. The curry puff skins were fried at 180 °C for 2, 3, 5, and 7 min. Before that, the frozen curry puff skins were thawed at room temperature for 2 h. The changes in colour, moisture, and texture were measured as indicators to Maillard reaction. Frozen curry puff skins reduced the level ofacrylamide and HMF to 70 % and 50 %, respectively after 28 days of storage. The colour development of frozen curry puff skins demonstrated less vividness as the storage time increased as compared to fresh curry puff skins. The fresh and frozencurry puff skins exhibited similar firmness and texture profile with no significant differences during the 4 weeks of storage.
In silico molecular modelling and docking of allicin, epigallocatechin-3-gallate and gingerol against colon cancer cell proteins
Asita Elengoe, Elina Sebestian
APJMBB 28 (4): 51-67
Article DOI:
Click here to download [PDF] [Supplementary Data]
Globally, colon cancer is the second most common cancer among men and women. There is an urgent need to search for a cure for colon cancer. Phytocomponents have shown to exhibit chemoprevention and chemotherapeutic effects related to colon cancer. Thus, phytocomponents can be used as the lead for new drug discovery. Computational biology approaches such as protein modelling and docking have helped in designing substrate-based drugs. In this study, three dimensional (3-D) models of tumour protein (p53), adenomatous polyposis coli (APC) and epidermal growth factor receptor (EGFR) were built using SWISS-MODEL; and their interaction with allicin, epigallocatechin-3-gallate and gingerol through blind docking were evaluated using BSP-SLIM server. These three target proteins are from colon cancer. Physiochemical characters of protein models were assessed through ExPASy’s ProtParam tool. Moreover, the protein structures were validated using PROCHECK, ProQ, ERRAT and VERIFY 3D servers. The protein models’ scores were within normal range. It also showed that the protein models were stable to proceed with the docking approach. Finally, the protein structures (target proteins) were docked successfully with allicin, epigallocatechin-3-gallate and gingerol (phytocomponent). The protein models had a strong interaction with the phytocomponents due to their good binding scores. The best docking scores of the protein-phytocomponent complexes (p53-allicin, APC-Epigallocatechin-3-Gallate and EGFR-gingerol) were 4.968, 6.490, and 6.034, respectively. Protein p53 had the strongest interaction with allicin due to its lowest binding score among all the protein-plant compound complexes. Thus, the results of this study can be used to design and develop a more powerful structure-based drug.
Bioremoval of Acephate by biofilm-forming Enterobacter cloacae – VITDAJ8 in a vertical packed bed biofilm bioreactor
Dipti Mayee Dash, Ankita Itusha, Jabez Osborne W
APJMBB 28 (4): 68-80
Article DOI:
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Organophosphorus (OP) pesticides are widely used due to their specific chemical properties and broad spectrum insecticidal activity. Accumulation of OP residues in various targeted and non-targeted environmental sites have increased the risk of biomagnifications thereby rendering serious threat to human health and the environment. Hence, the present study is primarily focused on the effective removal of Acephate (ACP) from aqueous solution. Bacteria isolated from the agricultural field and tannery effluent were screened based on their tolerance against two OP pesticides. The pesticide-resistant strains were further screened for biofilm formation and EPS production under ACP stress. Isolate VITDAJ8 exhibited highest pesticide tolerance against ACP, excellent biofilm formation and EPS production ability. Molecular characterization of VITDAJ8 by 16S rRNA gene sequencing revealed the isolate to be the closest neighbor of Enterobacter cloacae (Accession no. KP305912). A bench-scale vertical packed bed biofilm bioreactor was designed for the bioremoval of OP pesticides. Groundnut shells with a bed height of 10 cm were used as substrate with biofilm formed by VITDAJ8. The biosorption of ACP from the aqueous solution was monitored using UV-Vis spectrophotometer and HPLC and maximum ACP bioremoval up to 86% was achieved within 90 minutes of reaction time. Removal of ACP using groundnut shells without biofilm support was comparatively less efficient. The equilibrium adsorption study revealed the process to be the best fit for Langmuir isotherm modeling with maximum adsorption up to 235 mg L-1 and the kinetics of biosorption was found to be fitting well with the second-order kinetic model.
Heavy metal phytoremediation: Potential and advancement
Pushpa Chaudhary Tomar, Shilpa Samir Chapadgaonkar, Varsha Panchal, Arpita Ghosh
APJMBB 28 (4): 81-93
Article DOI:
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Industrial activities lead to the release of different types of toxic metals into the environment.  Phytoremediation has been established as one of the environmental-friendly and economical processes that have the potential for the remediation of industrial waste. Phytoremediation is used to extract metals from industrial effluents using ex-situ and in-situ treatments. Also, phytoremediation may be used to reclaim the polluted land resource for agricultural purposes. Moreover, this also prevents the bioaccumulation and biomagnification of xenobiotics from farming activities if carried out from polluted land. Phyto-mining can be done to recover and reuse the heavy metals from plant tissues after phytoremediation by plants. This study aimed to give a comprehensive review of recent research work in heavy metal phytoremediation.
MTHFR rs1801133 polymorphism in Bangladeshi population – its prevalence and detection
Sumaiya Farah Khan, Mahmuda Akter, Shayan Shahriar, MD Amzad Hossain, Abu Ashfaqur Sajib
APJMBB 28 (4): 94-101
Article DOI:
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Methylenetetrahydrofolate reductase (MTHFR) has been reported as a key enzyme not only for intracellular folate homeostasis but also for metabolism. A particular variant (G677A) leads to an altered amino acid, which ultimately causes decreased enzyme activity and may modulate the risk of causing several chronic diseases. The purpose of this study was to detect the pervasiveness of this variant MTHFR rs1801133 G677A in the Bangladeshi population. We applied allele-specific polymerase chain reaction (AS-PCR) to determine the genotypes at the rs1801133 in the Bangladeshi population. We performed targeted sequencing of the AS-PCR product of randomly selected samples. Out of the 181 Bangladeshi individuals, 71.8% had homozygous 677GG genotype, while 28.2% comprised of heterozygous 677GA genotype. No individual with the homozygous 677AA genotype was found in this representative Bangladeshi population. The 677G alleles had higher frequency (0.856) compared to 677A alleles (0.144) at the rs1801133 locus. Though the more risky homozygous 677AA genotype at the rs1801133 locus is absent in the Bangladeshi population, further association studies can be performed to identify the role of MTHFR mutation in the susceptibility to different multifactorial diseases.
Antigenic outer membrane proteins prediction of Pasteurella multocida serotype B:2
Farahani Muhammad Azam, Mohd. Zamri Saad, Raha Abdul Rahim, Pramote Chumnanpoen, Teerasak E-kobon, Sarah Othman
APJMBB 28 (4): 102-116
Article DOI:
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Outer membrane proteins (OMPs) are one of the prominent virulence factor or immunogenic element of Pasteurella multocidawhich are responsible for eliciting immune responses in multiple infected hosts. Identification of these proteins allows researchers to target OMPs to be manipulated as a vaccine against bacterial infection. Precise and rapid bioinformatics tools allow researchers to perform in silico analysis to extract putative OMPs from the genome information. In this study, we have successfully identified 105 putative OMPs of P. multocida subsp. multocida strain PMTB2.1 through computational prediction tools including a subcellular localisation predictor, PSORTb v3.0 followed by a lipoprotein predictor, LipoP 1.0 and a β-barrel transmembrane protein predictor, BOMP for sub-classification of the OMPs into 53 integral and 52 peripheral OMPs of this strain. The manipulation of antigenic epitope predictors and the antigenicity score filtering identified nine putative antigenic OMPs. These putative predicted antigenic OMPs of this pathogen will provide crucial initial guidance for the experimental identification and selection of antigenic protein(s) for the development of future haemorrhagic septicaemia (HS) vaccine.
Effects of maltodextrin concentration and inlet temperature on the physicochemical properties of spray-dried kuini powder
Yu Von Germaine Chng, Lee Sin Chang, Liew Phing Pui
APJMBB 28 (4): 117-131
Article DOI:
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Kuini (Mangifera odorata) is known for its intense flavour, attractive colour, rich fibre content and antioxidant properties. However, its short shelf life (bruise within a week) limits the availability of this fruit. Hence, spray drying was proposed to transform this fruit into powder to increase its application. The maltodextrin concentration of 5–20% (w/w) and inlet temperatures of 140–180°C were applied to produce kuini powders. The effect of maltodextrin concentrations at constant inlet temperature (160°C) showed that a higher amount of maltodextrin (20% w/w) produced powder with a low water activity (0.16 ± 0.01 Aw), moisture content (2.16 ± 0.60%) and hygroscopicity (20.45 ± 0.60 g/100g) but the orange colour of the powder turned pale. The kuini powder was further spray dried at different inlet temperatures at constant optimal maltodextrin concentration (20% w/w). The kuini powder that was spray dried at 160°C had the optimal properties: low water activity (0.17 ± 0.01 Aw) and hygroscopicity (22.12 ± 0.09 g/100g) with high water solubility index, WSI (79.90 ± 1.85%) and wettability (279 ± 8 s), indicating the powder can be applied as a food ingredient with high solubility. The optimal spray-drying condition for the production of kuini powder was 20% (w/w) maltodextrin concentration at 160°C inlet temperature to have the highest yield of 43.08 ± 2.77%. Eventhough the reconstituted kuini powder had lower viscosity, colour and β-carotene content than kuini juice, this study showed the feasibility of production of kuini powder using spray drying operation which may broaden its application in the food industry. 
Pre-treatment effects against the diclofenac-induced toxicity by the aqueous leaf extract of Madhuca longifolia on female Wistar albino rats for 10 and 15 days
Jerine Peter Simon, Sibi Karthik Saravanan, Jayashree Sathiyanarayanan, Radhakrishnan Vidya, Usha Kumari, Sabina Evan Prince
APJMBB 28 (4): 132-143
Article DOI:
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Diclofenac is used to treat rheumatism disorders, which are associated with the damages of renal, gastric and hepatic organs. Diclofenac is a pharmaceutical drug that is known to induce toxicity on its overdosage and long-term usage. Madhuca longifolia is known to have antioxidant, anti-inflammatory and anti-ulcer activity. It is an evergreen tree that is reported to have many ethnomedicinal uses. The other properties of Madhuca longifolia include anti-diabetic, analgesic and anti-microbial activities. Our study aims to evaluate the pre-treatment activity against the diclofenac-induced toxicity by the Madhuca longifolia aqueous leaf extract in Wistar albino rats for 10 and 15 days. Rats were divided as Group-I: Normal control, Group-II: Diclofenac on the last two days, Group-III and group-IIIa: Diclofenac + Aqueous Leaf Extract of Madhuca longifolia, Group-IV and group-IVb: Diclofenac + Silymarin, Group -V and group-Va: Aqueous Madhuca longifolia leaf extract. After the sacrifice, the rats were studied for antioxidant assay, renal enzyme markers, liver enzyme markers, and histopathological analysis of the kidney, stomach, intestine, and liver. As a result, we could identify that Madhuca longifolia has reduced the toxic changes in rats caused by diclofenac.


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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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