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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: https://doi.org/10.35118/apjmbb.2020.028.1.01
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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: https://doi.org/10.35118/apjmbb.2020.028.1.02
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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: https://doi.org/10.35118/apjmbb.2020.028.1.03
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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: https://doi.org/10.35118/apjmbb.2020.028.1.04
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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: https://doi.org/10.35118/apjmbb.2020.028.1.05
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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: https://doi.org/10.35118/apjmbb.2020.028.1.06
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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: https://doi.org/10.35118/apjmbb.2020.028.1.07
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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: https://doi.org/10.35118/apjmbb.2020.028.1.08
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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: https://doi.org/10.35118/apjmbb.2020.028.1.09
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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: https://doi.org/10.35118/apjmbb.2020.028.1.10
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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: https://doi.org/10.35118/apjmbb.2020.028.1.11
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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: https://doi.org/10.35118/apjmbb.2020.028.2.01
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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: https://doi.org/10.35118/apjmbb.2020.028.2.02
Click here to download [PDF] [Supplementary Data]
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: https://doi.org/10.35118/apjmbb.2020.028.2.03
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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: https://doi.org/10.35118/apjmbb.2020.028.2.04
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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: https://doi.org/10.35118/apjmbb.2020.028.2.05
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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: https://doi.org/10.35118/apjmbb.2020.028.2.06
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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: https://doi.org/10.35118/apjmbb.2020.028.2.07
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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: https://doi.org/10.35118/apjmbb.2020.028.2.08
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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: https://doi.org/10.35118/apjmbb.2020.028.2.09
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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: https://doi.org/10.35118/apjmbb.2020.028.2.10
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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.

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