Volume 33(1); 2025

Comparative analysis of microbial diversity in various kombucha starter cultures in Malaysia using the random amplified polymorphic DNA (RAPD) approach

Nur Syafiqah Syahmimie Augustine, Eric Tzyy Jiann Chong, Clemente Michael Vui Ling Wong, Nurul Farhana Nasir, Shaiful Adzni Sharifuddin, Noorjahan Banu Alitheen, Nurul Elyani Mohamad

APJMBB 33(1): 1-10

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.01

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Kombucha is sweetened black tea fermented with symbiotic culture of bacteria and yeasts (SCOBY) and has been widely consumed for its purported health benefits. The microbial consortium in kombucha is dominated by yeasts such as Brettanomyces sp. and Zygosaccharomyces sp., as well as acetic acid bacteria (AAB), Komagataeibacter sp. and Acetobacter sp. However, the source of SCOBY and substrates used may affect the microbial diversity as well as the biochemical, and flavour of the kombucha. Identifying the microbial population is important as the use of undefined starter cultures may lead to variable metabolite production and increase the risk of food pathogen contamination which can pose harm to human health. This study aims to isolate the microbes from kombucha in Malaysia for the future development of a new starter culture with specific species for the safe consumption of kombucha. Briefly, a total of 100 colonies were isolated from nine kombucha starter cultures with selective culture plates. Differences in colony morphology were observed based on their colours, surface texture, elevation, and margin. Their phenotypic morphology and genetic diversity were screened using microscopic examination, coupled with a catalase test and random amplified polymorphic DNA (RAPD), respectively. Based on the data, 51% of the isolates showed yeasts morphology under microscopic examination, while the rest were bacteria. Additionally, 55.79% of the isolates showed distinct banding profile patterns in the RAPD assessments. In conclusion, the data of this study shows that there is a diverse microbial consortium in different starter cultures of kombucha from Malaysia, mainly predominated by Gram-negative AAB and Gram-positive yeasts. Identification at the species level is to be conducted in the future. By understanding the microbiota diversity in kombucha, it contributes to the development and production of a safe functional drink.

The smiting of Streptococcus mitis and Streptococcus oralis: A novel antimicrobial proposition

Moad Khalifa, Wei Cun See Too, Ai Lan Chew, Ling Ling Few

APJMBB 33(1): 11-18

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.02

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S. mitis and S. oralis are typically found in the oral cavity of humans, but they have the potential to become harmful pathogens that cause infective endocarditis and bacteraemia, especially in individuals with weakened immune systems. Antimicrobial resistance (AMR) in bacteria is a serious issue that poses a significant threat to public health, and if appropriate measures are not taken, it could result in an estimated ten million deaths annually by 2050. S. mitis group of bacteria has demonstrated resistance to various antibiotics, highlighting the urgent need for novel antimicrobial strategies. Research has shown that inhibitors of human choline kinase can impede the activity of choline kinase (ChoK) and inhibit the growth of S. pneumoniae. ChoK, encoded by the licA gene of Streptococcus species, is responsible for producing phosphorylcholine, a precursor for lipoteichoic acid and cell wall teichoic acid. Due to the significant homology of the licA gene and protein sequences of S. mitis, S. oralis to S. pneumoniae, we propose that choline kinase inhibitors (ChoKIs), which have been effective against S. pneumoniae, have a promising prospect in inhibiting S. mitis and S. oralis. In summary, the development of novel antimicrobial strategies is critical to combat the spread of AMR in S. mitis and S. oralis. The high homology between the licA gene and protein sequences of these bacteria presents a promising opportunity for the development of ChoKIs to "smite" S. mitis and S. oralis and potentially mitigate the high mortality rates associated with AMR.

Role of Socs1 variants and Socs3 expression levels in interleukins production in asthma

Hanya Bachay Makttoof, Shereen Al-Ali, Hadi Lazim Alabadi

APJMBB 33(1): 19-26

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.03

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Asthma is a heterogenic chronic inflammation of the lower respiratory tract. Asthma is highly prevalent in Basrah city. This study aimed to investigate the effect of SOCS (SOCS1 & SOCS3) at the molecular level on the production of several cytokines (IL-6, IL-17 and IL-31) in asthma. The current study included a total of 80 participants, 54 asthmatic patients and 26 heathy controls. The levels of IL-6, IL-17 and IL-31 were estimated by ELISA while the SOCS1 gene polymorphism -1478 CA>del (rs33989964) was detected by RFLP-PCR. In addition, SOCS3 expression was estimated by qRT-PCR. The findings shown that the genotype del/del was significantly associated with asthma in the codominant model while CA/CA and CA/del genotype were significantly associated with asthma in the recessive model. The CA/del genotype was significantly associated with asthma in the over dominant model. The expression of SOCS3 was elevated non-significantly in asthma patients (4.28±1.41) while healthy control groups was 1. The highest level of SOCS3 expression was found in association with the CA/del genotype. The expression of SOCS3 did not have any effect on IL-6 level, while it had a slight effect on IL-17 in the CA/del genotype. The expression of SOCS3 has a clear effect on IL-31 in all SOCS1 genotypes. In conclusion, the heterotypic genotype CA/del plays important role in both the expression of SOCS3 and the production of IL-31 during asthma. 

Identification of putative bioremediation-related genes in purple non-sulfur bacteria isolated from a contaminated river in Malaysia

Oi Min Xian, Siti Halimah Hasmoni

APJMBB 33(1): 27-36

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.04

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Purple non-sulfur bacteria (PNSB) is a photosynthetic bacterium that can grow under different environmental conditions. These bacteria have been manipulated to develop biotechnological applications, including bioremediation and improving soil conditions and plant growth. In Malaysia, studies on nitrogen fixation and heavy metal removal capability of locally isolated PNSB are limited. Therefore, this study aimed to identify nitrogenase-related genes and heavy metal resistance genes in a strain of PNSB isolated from a locally contaminated river. Extracted genomic DNA was used in polymerase chain reaction (PCR) to detect target genes using specific primer pairs. The sequences were analyzed via Blastn and UniProt Blastx search to determine similar proteins and conserved sequence regions. Analysis of the amplified pufM gene identified Rhodopseudomonas sp. strain SBL as a purple phototrophic bacterial isolate. The nifH gene encoding for a molybdenum-nitrogenase was amplified for nitrogenase-related genes, indicating the role of Rhodopseudomonas sp. strain SBL in nitrogen fixation. Putative fragments of cueO and copA genes were detected in the genome of this isolate. No amplification of czcD and pbrA genes from this isolate was observed. Findings from this study showed the potential of Rhodopseudomonas sp. strain SBL as a nitrogen biofertilizer and a bioremediation agent in heavy metal contaminated agricultural land.

Mitigation of oxidative stress and testicular damage by Barringtonia racemosa aqueous extract in streptozotocin-nicotinamide induced diabetic rats

Nur Ahfizah Abdul Hadi, Siti Arffah Kamarulzaman, Siti Hajar Adam, Nik Nasihah Nik Ramli

APJMBB 33(1): 37-44

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.05

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Diabetes mellitus is a chronic metabolic disorder associated with various complications which include derangement in male reproductive functions. Barringtonia racemosa, has been studied widely as a complementary intervention in managing diabetes mellitus. However, the effect of B. racemosa on the male reproductive system is yet to be elucidated. Therefore, this study aimed to investigate the potential protective role of B. racemosa on male reproductive functions in type 2 diabetes mellitus (T2DM). The diabetic rats were administered 250 and 500 mg/kg/day body weight of B. racemosa aqueous extracts (BRAE) for 42 days. Fasting blood glucose was measured at the end of the experimental duration. Following sacrifice, the testes sections were examined for histopathological changes, and sperm count was assessed for further evaluation. Additionally, the testes homogenates were analyzed to determine antioxidant activity levels. Administration of BRAE ameliorated fasting blood glucose levels reduced structural damage to the testes, and improved sperm count. These findings were further supported by decreased lipid peroxidation and enhanced antioxidant enzyme activities in the testes of diabetic rats treated with BRAE. This study underscores the potential of BRAE, particularly at 250 mg/kg, as a promising candidate for managing diabetes-induced male infertility.

Structural and proteomic insights into the antibiofilm efficacy of DMSO on Corynebacterium pseudotuberculosis

Anati Abd Rashid Syaida, Mohd Izwan Mohammad Yusof, Faez Firdaus Abdullah Jesse, Nazia Abdul Majid, Mohd Fakharul Zaman Raja Yahya

APJMBB 33(1): 45-58

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.06

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Corynebacterium pseudotuberculosis is an intracellular pathogen particularly responsible for causing caseous lymphadenitis in sheep and goats, resulting in substantial economic losses. Dimethyl sulfoxide (DMSO) has been shown to inhibit the formation of C. pseudotuberculosis biofilm. However, the molecular mechanisms underlying the antibiofilm efficacy of DMSO against C. pseudotuberculosis biofilm remain poorly understood. The present study was conducted to evaluate the inhibitory effects of DMSO on C. pseudotuberculosis biofilm and to determine the structural and proteomic changes in the biofilm following DMSO treatment. Pellicle assays, biomass assays, viability assays, FTIR spectroscopy, and FESEM were used to assess biofilm formation, biomass, viability, biochemical composition, and morphology of C. pseudotuberculosis, respectively. Additionally, one-dimensional SDS-PAGE in combination with tandem mass spectrometry and bioinformatics were performed to identify proteins expressed in C. pseudotuberculosis biofilm following treatment with 50% (v/v) DMSO. Results showed that DMSO treatment significantly inhibited the pellicle formation, biomass, and viability of C. pseudotuberculosis biofilm. Biochemical and morphological changes in C. pseudotuberculosis were observed following treatment with 50% (v/v) DMSO. 1D SDS-PAGE revealed differential protein expression in the range between 33.7 kDa and 150 kDa following treatment with 50% (v/v) DMSO. A total of 387 proteins were differentially expressed between the control and 50% (v/v) DMSO-treated biofilm. Analysis of the protein-protein interaction network of proteins with differential expression revealed that 50% (v/v) DMSO had an impact on 10 biological processes and 18 hub proteins. It is postulated that DMSO may inhibit C. pseudotuberculosis biofilm by modulating multiple biological pathways.

Optimization of calcium, magnesium and zinc ions in yeast nitrogen base media for bioethanol production using response surface method

Safri Ishmayana, Sheila Farahdika, Debora Tamaris Horasio, Agus Safari, Muhammad Fadhlillah, Ukun M. S. Soedjanaatmadja, Retna Putri Fauzia, Fajriana Shafira Nurrusyda

APJMBB 33(1): 59-68

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.07

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The rapid decline in fossil fuel availability has driven a high demand for alternative fuels, including bioethanol. Bioethanol produced from starchy, sugary, or lignocellulosic materials through hydrolysis and fermentation by microorganisms, especially Saccharomyces cerevisiae. During fermentation, the yeast cell exposed to various stress factors that can reduce the ethanol productivity. One of the means to improve yeast cell tolerance against various stress factors is by supplementing fermentation media with metal ions. Metal ions have important role to increase the rate of sugar conversion to ethanol, cell viability and stress tolerance, and also the degree of final ethanol yield. This study aimed to determine the optimum concentrations of metal ions (calcium, magnesium, and zinc) in yeast nitrogen base (YNB) media for achieving the highest ethanol yield using S. cerevisiae Pinnacle S. The optimum condition was determined using response surface method – central composite design (RSM-CCD) approach. The results of this study indicate that the optimum concentrations of calcium, magnesium, and zinc ions for bioethanol production in YNB media are 26.36 mg/L, 31.82 mg/L, and 68.64 mg/L, respectively. Under these conditions, the maximum ethanol yield achieved was 0.197 g/g, representing a 4.8% increase compared to the central point (0.188 g/g), which was considered the baseline result.

Association between asthma and the tumor necrosis factor alpha-308 gene polymorphism in Iraqi patient

Rana Talib Mohsen, Wafaa Hussien Habeeb, Thaer Abdullah Hasan, Al-Taee Haneen Z

APJMBB 33(1): 69-76

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.08

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TNF-α has significant physiological effects on airway remodeling and inflammation. When asthmatic patients are exposed to allergens, peripheral blood monocytes and macrophages produce more TNF-α. In asthmatic individuals, TNF-α is the source of airway inflammation, late-phase airway hyperresponsiveness, and smooth muscle activation. An examination of the impacts of genetic variations in TNF-α-308G/A, as well as genotypes and alleles associated with asthma, has been conducted using molecular and immunological methods. A study involving 50 asthma patients (19 females and 31 males) and 40 healthy individuals (20 females and 20 males) demonstrated a correlation between polymorphisms and the likelihood of having asthma. The TNF-α-308G/A single nucleotide polymorphism (SNP-PCR) showed a significantly higher prevalence (P < 0.05) among asthma patients. The findings show that, compared to the control group, asthma patients had considerably higher levels of TNF-α-308G/A alleles and GA heterozygotes (P<0.05). This suggests that gene promoter polymorphisms may impact asthma sensitivity to TNF-α-308G/A.

In-silico analysis of Spike and envelope protein mutations in Omicron variants from Indonesia and high-case countries

Rahmadi Dimas Pratama, Erlia Narulita, Riska Ayu Febrianti, Aditya Kurniawan

APJMBB 33(1): 77-86

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.09

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Coronaviruses have an envelope made up of four main structural proteins, namely Spike protein that has a major role in binding and tends to undergo mutations. The purpose of this study was to analyze mutations that occur in the nucleotide sequence of the Indonesian Omicron variant Spike protein, which will be compared with the results of the analysis of Omicron variant mutations from 5 countries with the highest cases. The study began with a search for the sequence of the gene coding for the SARS-CoV-2 protein that had mutations from Indonesian patient isolates on the Global Initiative on Sharing All Influenza Data (GISAID) website. The obtained nucleotide sequence was then analyzed to determine the location of the mutations. The results of the sample analysis from 5 countries showed a composition of mutations that lead to decreased protein stability (11 mutations), 2 neutral mutations and mutations that cause increased stability (4 mutations). Mutations that cause decreased stability namely, G339D, K417N, N440K, T478K, E484A, Q498R, Y505H, D405N, R408S, L452R, and F486V, all cause disease. Mutations that reduce stability are G339D, S371L, K417N, N440K, G446S, T478K, E484A, Q493R, G496S, Q498R, and Y505H, with each mutation causing an increase in virulence. Mutations that increase stability namely, S373P, S375F, and N501Y. all three led to an increase in virulence. The results showed that the Omicron variant from Indonesia has higher stability and virulence then the other variants from other countries.

Assessment of genetic variability among Stevia rebaudiana Bertoni accessions cultivated in Malaysia using microsatellites

Pedram Kashiani, Hann Ling Wong

APJMBB 33(1): 87-95

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.10

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Stevia rebaudiana Bertoni (hereinafter “stevia”) is a herbaceous perennial plant of the Asteraceae family. Originating from Paraguay, stevia is considered a temperate, self-incompatible, insect-pollinated species. Recent sugar shortages in Malaysia have increased interest in stevia as an alternative. The stevia accessions available in Malaysia today are limited in number and perform poorly, emphasizing the necessity of varietal improvement. Assessment of genetic variability, diversity and intraspecific relationships is an essential step for such breeding efforts. This study aimed to reveal the genetic diversity among 21 stevia accessions, namely SRBA-1 to SRBA-21, collected from different origins utilizing 52 microsatellite DNA markers, and to identify potential markers for further marker-trait association mapping investigations. Results showed that 42 pairs of microsatellite primers (82.6%) produced polymorphic and reproducible bands. Average number of polymorphic bands ranged from 2 to 11, with an average of 5.1 alleles per locus. The total genetic diversity detected among the accessions ranged from 0.14 to 0.81, with a mean value of 0.58. Expected heterozygosity (He) and observed heterozygosity (Ho) (with mean values of 0.52 and 0.86, respectively) showed that almost all markers had deviation (Ho>He) from Hardy–Weinberg equilibrium (HWE). This result confirmed the highly outcrossing nature of this plant. The accessions evaluated revealed a wide range of variations in dry leaf weight, stevioside, rebaudioside A, rebaudioside C and total steviol glycosides. Eleven microsatellite markers with significant associations with the phenotypic traits were identified. Biosynthesis of steviol glycosides were found to be associated with the microsatellite markers SUGMS-23, SUGMS-26, SUGMS-28 and SUGMS-35. These findings can be utilised for developing marker-assisted selection procedures in stevia toward production of superior varieties for tropical environments.

Optimizing E. coli production of oil palm pla1-3 lipase through recombinant technology

Fatin Fathiah Safiudin, Chee Fah Wong, Zatty Syamimi Adura Mat Said, Ummirul Mukminin Kahar, Iffah Izzati Zakaria, Nurniwalis Abdul Wahab

APJMBB 33(1): 96-107

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.11

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The mesocarp of Elaeis guineensis is blessed with an abundance of oil proven to be graced by the presence of naturally occurring diverse family of lipase, many of which have yet to be studied.  These lipases influenced the innate oil quality while also being responsible for other biochemical reactions in the fruit. To accurately determine the capabilities of these lipase species, single protein isolation was necessary. PLA1-3; a potentially important oil palm lipase, was selected for preliminary expression and purification to facilitate later functional study. The need for targeted, gene-specific analysis had influenced the employment of recombinant protein strategy in this research. PLA1-3 of oil palm mesocarp was cloned in pET28a(+) and expressed as fusion His-tagged protein with E. coli as host. It was revealed that PLA1-3 was best expressed in Rosetta 2 (DE3) pLysS with 2 mM of IPTG at 30°C. PLA1-3 was able to be partially purified with 210.1 U/mg of specific lipase activity. PLA1-3 evidently exhibited biocatalytic property which promised a prospective industrial application in the near future.

Purification and characterisation of Bile-salt activated lipase (BSAL) enzyme from bovine (Bos taurus) pancreas

Nur Hidayah Reshidan, Norliza Bahtiar Affendy, Mohd Anuar Jonet, Dhilia Uddie Lamasudin, Zaidah Zainal Ariffin, Mazatulikhma Mat Zain, Umi Marshida Abd Hamid

APJMBB 33(1): 108-118

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.12

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Bile salt-activated lipase (BSAL) is one of the pancreatic lipases that plays a critical role in the digestion and absorption of dietary fats. Objective: This study aimed to purify bile salt-activated lipase (BSAL) from bovine (Bos taurus) pancreatic tissue. Methods: Bovine pancreas was freshly collected from Abattoir Complex, Department of Veterinary Services, Shah Alam, Malaysia. The fats were removed by trimming grossly visible fat, and extraction was performed using organic solvents. The BSAL was purified by anion exchange chromatography and sent for protein identification by liquid chromatography-mass spectrometry (LC-MS/MS). Successful purification of bovine BSAL was visualised as a single protein band on sodium dodecyl sulphate-polyacrylamide (SDS) gel, which LC-MS/MS confirmed as a bovine BSAL (Accession number – P30122) with a molecular mass of 65.12 kDa and calc pI of 5.57. Peptide identification based on the MS spectrum found 200 predictive peptides, ten sequences with bovine BSAL peptide characteristics. The selected predictive peptide sequences have a molecular mass of 1104.60 - 3378.94 Da with Qvality q-value greater than 0.01 and XCorr Sequest HT value ranging from 2.6 to 6.8. The specific lipolytic enzyme activity of bovine BSAL was comparable with the positive control, as measured using lipase assay. In conclusion, the results of this study indicate the effectiveness of bovine BSAL purification by anion exchange chromatography from fresh pancreatic tissue and may have the potential for further Halal pharmaceuticals and medical applications. 

Optimising callus induction in Kappaphycus alvarezii via micropropagation techniques under different LED light conditions

Muhammad Asyraf Abd Latip, Siti Noor Ain Hamzah, Najihah Mohamad Nasir, Syed Mohamad Azim Syed Mahiyuddin

APJMBB 33(1): 119-124

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.13

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High demands of Kappaphycus alvarezii in markets require commercial scale of the seaweed’s cultivation and large amount of seedlings. One of the best ways to overcome seedling scarcity is by using seaweed micropropagation. This study was carried out to produce K. alvarezii type Giant Tambalang tissue culture by using micropropagation technique. The axenic explants were inoculated into petri dishes with plant agar and 1.5% agarose supplemented with Provasoli’s enriched seawater (PES). The explants were cultured at 25 °C ± 1°C under cool white LED light (Culture room 1(CR1)) and red and blue LED lights (Culture room 2 (CR2)) at 17.55 µmol m^-2s^-1 and at 20.28 µmol m^-2s^-1, respectively. After 6 days, the results showed that the method of treating the thallus by using 70% alcohol for 5 seconds in sterile artificial seawater is able to induced calluses for explants cultured in CR1 with rate of callus formation at 53.33% (plant agar) and 46.67% (agarose). Explants cultured in CR2 showed sign of calluses growth after 9 days with the rate of 56% (plant agar) and 76% (agarose). After 12 days, the callus percentage in both culture room increased significantly in CR1: 72% (plant agar) and 77.33% (agarose); and in CR2: 68% (plant agar) and 81.33% (agarose).  For this analysis, the optimal parameters for callus induction were observed when the explants were cultured on 1.5% agarose agar supplemented with PES at 25°C ± 1°C under a combination of red and blue LED lights at 20.28 µmol m^-2s^-1. Further experiments need to be carried out to be able to achieve significant results.

ASAP 1 genetic polymorphisms and tuberculosis: A comprehensive review of susceptibility markers

Ismail Ismail, Sukriyadi Adi, Muhammad Basri, Simunati Simunati, Nasrullah Nasrullah

APJMBB 33(1): 125-131

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.1.14

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ASAP1 polymorphisms have been investigated for their potential association with tuberculosis (TB) susceptibility in various populations. This review aimed to investigate the association between ASAP1 polymorphisms and susceptibility to TB in various populations. A systematic review of relevant studies was conducted, focusing on genetic association analysis and functional studies investigating ASAP1 expression and its role in TB pathogenesis. Data were drawn from various populations, including Russian, African, Chinese, Mongolian, and Tibetan groups, to assess genetic diversity in TB susceptibility. A genome-wide association study in Russia identified seven single nucleotide polymorphisms (SNPs) in ASAP1 associated with TB susceptibility. However, studies conducted in China's Xinjiang Muslim population yielded mixed results, with two of these SNPs significantly associated with TB risk. The A allele of rs4733781 was associated with increased TB risk (OR = 1.242, P = 0.046), while the G allele of rs1017281 was associated with decreased risk (OR = 0.792, P = 0.028). In contrast, studies in Mongolian, Tibetan, and Han Chinese populations found no significant association, suggesting potential ethnic and environmental influences. However, some individuals with ASAP1 SNPs (rs10956514, rs4733781, rs2033059, rs12680942, rs1017281, rs1469288, and rs17285138) tended to have a reduced risk of TB, although this difference was not statistically significant. The biological role of ASAP1 in dendritic cell migration suggests TB susceptibility and host-pathogen interactions; yet, functional validation remains incomplete. This review highlights gaps in current knowledge, emphasizing the need for large-scale multi-ethnic studies, functional genomics approaches, and epigenetic analyses to elucidate the role of ASAP1 in TB pathogenesis.

Volume 33(2); 2025

Prevalence of polyketide synthase (PKS) genotoxin among Escherichia coli isolated from clinical sources 

Abdullah Ayoub Kamel, Suhad Saad Mahmood

APJMBB 33(2): 1-7

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.01

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Although some variants of Escherichia coli are harmless and even advantageous, certain strains are pathogenic and can lead to severe illness, including the onset of colorectal cancer associated with E. coli’s colibactin genotoxin. This study aims to determine the prevalence of the ClbA gene (gene of polyketide synthase (PKS) genotoxin) among E. coli isolates collected from various clinical sources. In this study, 120 samples were collected from different sources, and only 63 of those samples were positively identified as E. coli by utilizing biochemical tests, selective media, and PCR technique using the UidA gene. The results showed that only 33 / 63 isolates were clbA+ E. coli, while 42/63 isolates were uidA+ E. coli by using conventional PCR. Moreover, q-PCR was conducted on three isolates from different sources to detect ClbA gene expression. The results showed that ClbA gene expression was higher in E. coli isolates from colorectal cancer samples, followed by E. coli taken from wounds and then E. coli taken from urine. This study has found that E. coli possesses the pks-associated ClbA gene in wound infections more frequently than other sources. This gene appears the least in stool specimens and an aberrantly high expression of the clbA isolated from the pks+ E. coli in colorectal cancer patients.

Lapatinib modulates Caco-2 intestinal monolayer permeability through ErbB1 inhibition-mediated tight junction alteration

Raja Nur Firzanah Syaza Raja Sharin, Jesmine Khan, Mohammad Johari Ibahim, Joanne Bowen, Wan Nor I’zzah Wan Mohamad Zain

APJMBB 33(2): 8-23

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.02

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Lapatinib (LAP), a dual inhibitor targeting ErbB1 and ErbB2 tyrosine kinases, demonstrates efficacy in treating ErbB2-positive breast cancer. However, its clinical use is hindered by the occurrence of associated diarrhoea. ErbB1 is expressed in the intestine; leading to the hypothesis that lapatinib inhibits normal ErbB1 function, causing diarrhoea. This study explores the potential role of ErbB1 inhibition in the mechanism of lapatinib-induced diarrhoea. Caco-2 intestinal monolayers were treated with LAP alone and in combination with recombinant epidermal growth factor (LAP+rEGF). The integrity of the Caco-2 monolayer was evaluated by measuring transepithelial electrical resistance (TEER) of the Caco-2 monolayer and Lucifer yellow paracellular transport. Changes in tight junction proteins (TJPs) claudin-1, occludin and ZO-1 were assessed through qPCR and immunofluorescence staining, while the expression levels of inflammatory cytokines TNF-α, IL-1β, and IL-6 were determined using qPCR. At 96 hours, LAP treatment resulted in a significant reduction in TEER compared to the untreated control monolayer (p < 0.05). Although not significantly different from the control group, the LAP group exhibited higher Lucifer yellow permeability. LAP suppressed mRNA and protein expression of TJPs, with cotreatment of rEGF mitigating LAP inhibition significantly (p < 0.05). No significant changes were observed in the mRNA expression levels of inflammatory cytokines in the LAP group. However, treatment with rEGF increased IL-6 mRNA expression significantly (p < 0.01). Lapatinib increased Caco-2 intestinal monolayer permeability and reduced tight junctions by inhibiting ErbB1, suggesting a mechanism of lapatinib-induced diarrhoea. 

Current photodynamic therapy and emerging synergies in oncology

Danesh Thangeswaran, Ding Sheng Chin, Venugopal Balakrishnan, Sasidharan Sreenivasan, Hung Wei Lai, Gee Jun Tye

APJMBB 33(2): 24-40

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.03

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Cancer treatment faces ongoing challenges owing to the diverse cancer types and drawbacks of traditional therapies, prompting the search for less invasive alternatives. Photodynamic therapy (PDT) is a non-intrusive approach that uses photosensitisers (PS) to accumulate in target cancer cells and induce cell death upon light activation. This review critically evaluates the applications of two progressive PS, aminolevulinic acid (ALA) and indocyanine green (ICG), in PDT. It synthesizes findings from extensive studies and clinical trials to assess the current landscape of PDT and its potential in combination therapies. Additionally, this review explores the feasibility and rationale for integrating PDT with alternative therapies, such as sonodynamic therapy (SDT), radiodynamic therapy (RDT), photopheresis, photothermal therapy (PTT), and LED-phototherapy, to enhance treatment efficacy. ALA and ICG demonstrate significant potential for PDT owing to their selective accumulation in cancer cells and favorable safety profiles. Clinical trials have shown promising results, indicating PDT’s efficacy of PDT for various cancer types. Moreover, the combination of alternative therapies underscores PDT’s versatility and capacity for synergistic effects in cancer treatment. ALA and ICG are promising PS candidates that offer targeted cell death with minimal toxicity during PDT. The integration of PDT with alternative therapies presents a compelling avenue for enhancing the treatment outcomes and therapeutic potential in oncology. Further research and clinical validation are warranted to refine the full potential of PDT and its synergistic applications in cancer therapy.

Exploring termite-microbe symbiosis: mechanisms, evolution and biotechnological potential

Mardani Abdul Halim, Clemente Michael Wong Vui Ling, Kamarul Zaman Zarkasi, Homathevi Rahman, Nur Athirah Abdullah, Abdul Hafiz Ab Majid, Zarina Amin, Nurul Akmar Hussin

APJMBB 33(2): 41-53

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.04

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The symbiotic relationship between termites and their gut microbes is one of nature's most fascinating mutualistic interactions. Termites, renowned for their ability to decompose lignocellulosic materials such as wood, rely heavily on their gut microbiota, which includes bacteria, archaea, and protozoa, each playing specific roles in breaking down cellulose, hemicellulose, and lignin. This complex partnership not only enables termites to thrive in low-nutrient environments but also supports nitrogen metabolism and provides essential nutrients for their survival. The termite gut is highly compartmentalized, creating various microenvironments that support diverse microbial communities and facilitate metabolic processes, including fermentation in anoxic conditions. Shaped by evolutionary pressures, this symbiotic relationship has resulted in specialized adaptations that optimize lignocellulose degradation and nitrogen cycling. This review explores the mechanisms of termite-microbe symbiosis, their functional roles across different taxa, the structural and functional compartmentalization of the termite gut, and the evolutionary dynamics driving this mutualistic relationship. In addition, this review highlights the ecological and biotechnological significance of termite-microbe symbiosis, offering valuable insights into efficient lignocellulose degradation and nutrient recycling mechanisms that could inspire future applications in sustainable bioenergy and waste management.

Antimicrobial effects of oregano, Habbatus sauda and myrrh essential oils against bacteria causing nosocomial infections

Iman Batrisyia Hannani Mat Homsa, Khairiyah Murad, Maryam Al-Talib, Hassanain Al-Talib

APJMBB 33(2): 54-62

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.05

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Essential oils (EOs), including oregano, habbatus sauda, and myrrh, are increasingly coming into focus as new medicinal strategies against various organisms, including bacteria that cause nosocomial infections. The uncontrolled use of antibiotics is associated with multi-drug resistance. Therefore, it is crucial to find an alternative therapeutic approach to combat nosocomial infections. This study aims to investigate the antibacterial effects of oregano, habbatus sauda, and myrrh essential oils against the bacteria that cause nosocomial infections. The antimicrobial activity of the essential oils against methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, Escherichia coli and Staphylococcus epidermidis was investigated using the disc and well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by broth microdilution tests using essential oil concentrations between 100% and 0.19%. Oregano and habbatus sauda showed excellent inhibitory effects on nosocomial bacteria. However, myrrh was not effective against nosocomial bacteria. The MIC of oregano against MRSA, P. aeruginosa and S. epidermidis was 3.13%. The MIC of habbatus sauda against MRSA and S. epidermidis was 0.78%, while against P. aeruginosa and E. coli it was 50% and 12.5%, respectively. In conclusion, both oregano and habbatus sauda have excellent antimicrobial activity against nosocomial bacteria and are recommended as new alternative treatments for nosocomial infections.

Unlocking the molecular targets of ursolic acid for diabetic wound healing

Brigitta Amanda Maharani, K. Ariex Widyantara, Delicia Noviachen, Agustina Setiawati

APJMBB 33(2): 63-71

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.06

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Diabetic wounds, which account for 85% of non-traumatic amputations due to necrosis and gangrene, are a leading cause of hospital admissions among diabetic complications. These wounds require protective layers against infection and bioactive materials like ursolic acid (UA) to promote healing processes. This study investigates the molecular targets of UA through bioinformatics and in silico analysis to inhibit diabetic wounds. After obtaining an online database of the genes related to diabetic wound healing, the genes influenced by UA were identified using a Venn diagram. Subsequently, the top 10 target proteins associated with molecular pathways were scrutinized using CytoHubba to elucidate their roles. Finally, a molecular docking study was performed on TP53 and TNF for further investigation. UA orchestrates molecular cascades involving apoptosis and pyroptosis-related genes in many types of cells such as fibroblasts, keratinocytes, endothelial, and erythrocytes. This study is expected to promote the development of UA-containing natural products as wound healing agents for diabetic patients. This study may also support in vitro and in vivo studies of UA and UA-containing natural products in diabetic wound healing activities.

Investigation on the effects of siderite and biochar as mixed nutrient sources for bioremediation of synthetic groundwater Cr (VI) pollution

Shuqin Wang, Yuling Zhu, Baowei Hu, Mohamad Faiz Foong Abdullah, Zaidah Zainal Ariffin

APJMBB 33(2): 72-82

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.07

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Cr (VI) groundwater pollution poses a significant threat to both agricultural production and human health quality worldwide. Microbial-mediated Cr (VI) reduction offers an eco-friendly and cost-effective strategy. Although siderite and biochar have individually shown promise as electron donors for microbial remediation, their combined use remains underexplored. This study utilized siderite and biochar as mixed nutrient sources to construct bioremediation columns, investigating the microbial community's removal of Cr (VI) within the system. Results revealed that, at an influent Cr (VI) concentration of 0.6 mmol/L, the mixed nutrient source of siderite and biochar achieved a Cr (VI) removal rate ranging from 79.5% to 100%, significantly outperforming the siderite-only and biochar-only groups. XRD spectra confirmed the reduction of Cr (VI) to insoluble Cr (III) through both microbial and chemical pathways. The high microbiota diversity in the siderite and biochar columns, including Aquabacterium, Acidovorax, Reyranella, and Pseudoxanthomonas, was identified as a plausible explanation for the observed high removal rate of Cr (VI). These findings provide a theoretical basis for applying mixed nutrient sources, including siderite, in the removal of Cr (VI) from groundwater, highlighting the potential of this approach for eco-friendly and efficient remediation.

Identification of antimalarial drug candidates from natural products for Plasmodium falciparum lactate dehydrogenase (PfLDH) inhibition: In silico approach

Nursyahidatul Azwa Awang, Nurhainis Ogu Salim, Ernie Zuraida Ali

APJMBB 33(2): 83-93

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.08

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The increases of strains of malarial parasite resistant to existing medication treatment urges the look for antimalarials with novel methods of activity. Lactate dehydrogenase (LDH) is well-known as the potential target for the development of novel antimalarial drugs and later prevent the spread of the disease. Substrate binding site on LDH enzyme is an attractive and promising target for the development of inhibitors specific to malaria enzyme due to the distinctive cleft formed alongside the extended substrate specificity loop. In reaction to this, the purpose of the present study was to identify inhibitor from natural products which could be further optimized to become potential antimalarial drug candidates. Computational approaches which include molecular docking, drug-likeness and ADME prediction were used to screening the target inhibitors. The binding affinity of 13,21-Dihydroeurycomanone was found to be the highest with a value of -7.04 kcal/mol among the listed natural compounds. The result shows that this compound exhibited lower binding affinity than the reference ligand (BIH), however higher than oxamate showing a potential competitive binding. Besides, the docking result also suggested that 13,21-dihydroeurycomanone potentially promoted additional hydrogen-bond interactions with residue in the targeted binding site. Furthermore, the drug-like properties showed that 10 out of 13 compounds including 13,21-Dihydroeurycomanone, were acceptable that could be further investigated for designing of novel drug-like compounds for treatment of malaria. Taken together, this study successfully generated a list of potential inhibitors targeting substrate binding site to be developed as antimalarial drugs.

Rice (Oryza sativa) and its potential application in promoting wound healing: A narrative review

Vivi Noryati Ahmad, Nurhayati Mohamad Zain, Indah Mohd Amin

APJMBB 33(2): 94-106

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.09

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Wound healing stages are interlinked, with hemostasis initiating the process, followed by inflammation, proliferation, and remodeling. Chronic wounds exhibit prolonged inflammation and disrupted proliferation and remodeling stages, leading to impaired healing. This rise in chronic wounds reflects an increase in patient numbers and the need for prolonged treatment, highlighting the financial burden on healthcare systems and the economic impact on patients. Despite medical advances, wound healing poses challenges, necessitating ongoing research for novel therapies. Natural remedies like rice offer a promising avenue for wound management due to their antioxidant, anti-inflammatory, antimicrobial, regenerative and wound healing properties, potentially reducing healthcare costs and improving patient outcomes. Rice contributed to tissue repair properties, suggesting its integration into advanced wound healing strategies. This review emphasizes rice’s impact on wound healing across various contexts, including in vitro, in vivo, regenerative medicines as well as genetically modified rice. Research indicated rice’s efficacy in promoting healing while exhibits non-cytotoxic properties. Nonetheless, there is a lack of detailed in vitro studies on its effects on oral cells like human gingival fibroblasts and oral keratinocytes, and more in vivo research also is needed. This identified research gap presents direction for future inquiry, particularly within dental contexts. The potential of rice in advanced wound care and tissue regeneration represents a stimulating area for continued research. 

Identification and gene expression analysis of mating type (MAT) 1-1-1 gene in Fusarium oxysporum f.sp. cubense Tropical Race 4

Shin Huey Ang, Jia Xin Ong, Razak Terhem, Mohd Termizi Yusof, Mui Yun Wong, Tsutomu Arie, Noor Baity Saidi

APJMBB 33(2): 107-114

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.10

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Fusarium oxysporum f.sp. cubense Tropical Race 4 (Foc TR4) is a soil-borne fungal pathogen that impacts banana production. It is a member of the F. oxysporum species complex (FOSC), whose members are predominantly asexual. However, the FOSC members, including Foc TR4, retain mating type (MAT) loci, suggesting functions that are different from mating. This study identified and characterized the MAT1-1 locus in Foc TR4 and the expression of MAT1-1-1, the key gene in the MAT1-1 locus, during its interaction with susceptible bananas in vitro. The open reading frame of the identified MAT1-1-1 perfectly matched the reference sequence, and the corresponding MAT1-1-1 protein (367 amino acids) was predicted to localize to the nucleus, which matches its previously reported function as a transcriptional regulator. Based on the phylogenetic analysis of MAT1-1-1 protein sequences, Foc TR4 was grouped closely with other F. oxysporum formae speciales and clearly separated from other Fusarium species. The expression level of MAT1-1-1 was negligible in the absence of the host but was significantly induced when the host was present within a close range (at 2cm away) and at four to eight hours after being exposed to the host. These data reveal that the MAT1-1-1 of Foc TR4 is possibly associated with host sensing, allowing the pathogen to recognize specific signals from the host and activating genes required for directed hyphal growth and colonization. Overall, this study sheds light on the other potential roles of MAT genes, particularly MAT1-1-1 in Foc TR4.

Preparation of methanolic extract and silver nanoparticles from leaves of Iraqi Sesuvium portulacastrum plant and assessment of their antibacterial activity

Mustafa F. Hameed, Teif A. Najm, Bahaa Wathook Abdaljabar

APJMBB 33(2): 115-122

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.11

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Sesuvium portulacastrum (S. portulacastrum) is considered one of the most important plants, its cultivation has spread in Iraq recently, and it has great medical importance. Preparation and characterization of silver nanoparticles (AgNPs) and methanolic extract of S. portulacastrum leaves by microwave-assisted extraction (MAE) and comparison of their biological effects on some bacteria. In this study, microwave-assisted extraction was used to prepare silver nanoparticles and methanolic extract, and their antibacterial activity was tested. Silver nanoparticles were characterized by GC-MS, FT-IR, UV–visible, and Scanning Electron Microscopy (SEM). The microwave-assisted extraction method was devised to prove its efficiency in extraction. The composition of silver nanoparticles was proven through GC-Ms, IR, UV-visible, and SEM spectra. silver particles proved highly effective in inhibiting the growth and reproduction of bacteria of the type Escherichia coli (E. coli) compared with the methanolic extract. The current study did not prove any possible effectiveness against the growth and reproduction of bacteria of this type, Staphylococcus aureus (S. aureus). The study proved that microwave technology has excellent efficiency in terms of time, accuracy, and lower costs; silver nanoparticles proved to have more anti-growth activity than E. coli. This indicates the efficiency of these molecules towards microorganisms and their use as a future drug in treating the diseases caused by bacteria faster and better than the methanolic extract.

BAP combined with GA3 improve in vitro shoot multiplication for shoot tip and nodal explants of Rosa Damascena Mill 

Mansour A. Abohatem, Yousra Al-Qubati, Hanan Abohatem

APJMBB 33(2): 123-130

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.12

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Damask rose (Rosa damascene Mill) is one of the most valuable and oldest types of roses and an aromatic and medicinal plant. Since each of NAA and GA3 combined with BAP has a different effect on shoot regeneration, this study aimed to identify a suitable combination medium for shoot multiplication of Rosa damascene Mill. Induction and multiplication of shoots from both shoot tip and nodal explants were developed using various concentrations of 6-Benzylaminopurine (BAP) combined with 1-Naphthylacetic acid (NAA) or Gibberellic acid (GA3). The basal medium supplemented with BAP and GA3 showed the multiplication rate of 2 folds for both shoot tip and nodal explants compared to the medium supplemented with BAP and NAA. The combination medium of 1 mg/L BAP and 0.5 mg/L GA3 showed a maximum shoot number (4) and shoot length (4.6 cm) during the shoot multiplication period. In this study, the efficient role of the medium for BAP combined with GA3 was confirmed in the improved effectiveness of in vitro proliferated shoots protocol for Damask rose. This protocol is beneficial for the commercial micropropagation of in vitro rose plants.

Exploration of methanotroph bacteria in ponds with molecular approaches 

Ananda Tania Salsabila, Aninditia Sabdaningsih, Haeruddin

APJMBB 33(2): 131-137

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.13

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Semarang City as the capital city of Central Java Province is threatened by increasing the amount of greenhouse gas (GHG) emissions. The most study GHG is related to carbon dioxide emission, however, methane gas is one of the most dangerous GHG. Furthermore, the methanotroph bacteria in ponds are under-explored, therefore this study aims to isolate methanotroph bacteria and find species through molecular identification in ponds near Tirang Beach, Semarang. Isolation of bacteria initially used Nitrate Mineral Salt (NMS) media followed by specific media based on pMMO (Particulate Methane Monooxygenase) and sMMO (Soluble Methane Monooxygenase) activities, and molecular tests. The pMMO results were 0.013 to 0.105 mol/L, while the sMMO  obtained 0.230 to 0.399 mU/mg. Bacterial colonies obtained from NMS agar have an abundance of 3.29x10² CFU/mL. According to BLAST homology results on isolate T1.S.Al/3 has a similarity 98.81% with Methylophaga marina. This finding indicated that Methylophaga marina are methanotroph bacteria from its enzyme activities. 

Evaluation of the role of ERAP1, RUNX3, and HLA-C genes in Iraqi patients with psoriasis: A histopathological study

Widad Abed Jawad, Noor Mahmood Majeed

APJMBB 33(2): 138-143

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.14

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Psoriasis is a chronic disease caused by an overactive immune status, leading to the rapid multiplication of the skin cells and the formation of scaly, inflamed patches on various parts of the body. This study aims to determine the gene expression of ERAP1, RUNX3, and HLA-C in Iraqi patients with psoriasis. Seventy serum samples were collected from individuals attending Yarmouk Teaching Hospital in Baghdad, Iraq. The current study included two groups: Group 1 (G1) consisted of 50 patients with psoriasis, while Group 2 (G2) included 20 healthy individuals. Gene expression was assessed using quantitative polymerase chain reaction in both groups. Moreover, gene expression was analysed for both acute and chronic forms of psoriasis. The results showed that the expression of ERAP1 and HLA-C genes was increased in patients with psoriasis compared to G2. In contrast, the expression of RUNX3 gene was decreased in patients with psoriasis compared to G2. In addition, the expression of ERAP1 and HLA-C genes was significantly higher in the acute form of psoriasis compared to the chronic form and G2, while the expression of the RUNX3 gene was reduced in the acute form compared to the chronic form and G2. These findings suggest that the upregulation of ERAP1 and HLA-C genes and the downregulation of RUNX3 gene are indicative of psoriasis, particularly in its acute form. These evidence supports the potential involvement of these genes in the pathogenesis of psoriasis and highlights their distinct roles in acute and chronic cases of the disease.

Exploring the impacts of terminal mutations on the thermostability of Bacillus sp. L2 lipase

Siti Nor Hasmah Ishak, Khairina Alya Mohd Faizal, Mohd Ishtiaq Anasir, Noramirah Bukhari, Nur Aisyah Zulkifli Lubis, Mohd Radzman Mat Radzi, Adam Thean Chor Leow, Mohd Shukuri Mohamad Ali, Fairolniza Mohd Shariff

APJMBB 33(2): 144-153

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.15

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Protein engineering has been widely used to improve enzyme properties and make them appropriate for use as industrial biocatalysts. To study the effect of mutation at the N- and C-terminal of lipase, two double mutants (A8V/S385E and A8P/S385E) were generated by site-directed mutagenesis of L2 lipase from Bacillus sp. (wt-L2). The simultaneous mutations in the A8V/S385E and A8P/S385E resulted in significant changes in the lipase's properties compared to the wild-type (wt-L2). The mutants demonstrated increased thermostability compared to the wild-type. The melting temperature (T_m) analysis using circular dichroism revealed higher T_m values of 84.5 °C for A8P/S385E and 75.1 °C for A8V/S385E. This indicates that the enzyme can withstand higher temperatures before denaturation, a desirable trait in various industrial processes. Secondary structure analysis indicated alterations in the lipase structure caused by the simultaneous mutations. In summary, the simultaneous mutation at the C- and N-terminals had a multifaceted impact on the lipase, influencing its optimal temperature, thermostability, and structural characteristics. These findings provide insights into how specific genetic modifications can be employed to tailor the enzyme for improved performance in industrial applications.

Terminalia catappa ethanolic extract attenuates the growth of nosocomial pathogenic bacteria

Anis Syafiqah Norhisham, Kharul Azmi Mu’azzam Abdul Rahman, Nurul Aili Zakaria, Syarifah Ab Rashid, Mohd Fakharul Zaman Raja Yahya, Mohd Taufiq Mat Jalil

APJMBB 33(2): 154-167

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.16

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Nosocomial infections, or hospital-acquired illnesses (HAIs), are significant global health concerns. These infections reduce the efficiency of conventional antibiotics and contribute to the rise of multidrug-resistant strains, especially among ESKAPEE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Escherichia coli). ESKAPEE bacteria are highly virulent, resistant to antibiotics and frequently causing nosocomial infections. Hence, the present study aims to evaluate the efficacy of Terminalia catappa ethanolic extract (TCEE) against selected ESKAPEE strains. The antibacterial activity and efficacy of TCEE were determined using disc diffusion and broth microdilution assays. In vivo antibacterial activity of TCEE was assessed using a shake flask system to measure bacterial growth reduction, and a kill-time study was performed to evaluate the effectiveness of TCEE. In the disc diffusion assay, S. aureus was most susceptible to the TCEE with an inhibition zone diameter of 16.0 ± 1.7 mm, while E. coli showed the least susceptibility, with an inhibition zone diameter of 9.3 ± 0.6 mm. The MIC and MBC values revealed that the extract showed a bacteriostatic effect on all four test bacteria. Time-kill analysis showed that the killing effect of the extract was time- and dose-dependent, whereby a higher concentration of extract could kill bacterial cells at a shorter incubation period. The membrane cell leakage assay suggested that higher TCEE concentrations caused greater membrane cell disruption, as indicated by higher amounts of protein and nucleic acid in the medium. The present study suggests that TCEE could be a promising antimicrobial agent with a broad range of inhibitory effects. 

In-silico analysis of papain fibrinolytic mechanism from Carica papaya

Jonathan Suciono Purnomo, Dikson, Samuel Emmanuel Soentoro, Erika, Marcelia Sugata, Ariela Samantha, Reinhard Pinontoan

APJMBB 33(2): 168-182

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.17

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Thrombosis is an abnormal blood coagulation caused by imbalances in coagulation processes which may cause complications including cardiovascular diseases (CVDs). Current thrombolytic treatments are fast and effective but are expensive and risky, which prompts exploration of plant-based sources such as Carica papaya’s papain shown to have promising fibrinolytic capabilities. This study aims to elucidate papain's mechanism of action of fibrinolysis at a molecular scale through in-silico molecular docking and molecular dynamics (MD) analyses using AlphaFold3 and GROMACS respectively. Seven octameric fibrin peptides were predicted to have papain's cleavage targets, namely FibAα-1 and FibAα-2 from fibrin chain Aα, FibBβ-1, FibBβ-2, FibBβ-3, and FibBβ-4 from chain Bβ, and Fib𝛾-1 from chain 𝛾, all showing high binding affinity to papain, with dissociation constant (K_d) ranging from 7.6×10^-9 to 2.9×10^-7 M. MD simulations indicated stable docking positions, with confirmed low Root Mean Square Deviation (RMSD) and normalized Root Mean Square Fluctuation (RMSF_N) values. Most peptides, except FibAα-1, frequently approached the critical catalytic distance of 3.3 Å, suggesting their potential cleavability. This distance measures the gap between the sulfur atom of papain’s Cysteine-25 (SG) and the carboxyl carbon of the scissile peptide bond. Binding free energy (ΔG_bind) distributions showed lower binding energy is required for FibAα-2, FibBβ-2, FibBβ-3, and FibBβ-4 than for the other peptides. Notably, FibAα-2 and FibBβ-4, showed better docking and MD performance, suggesting they may be preferentially cleaved by papain. These findings underscore the potential of papain as a novel fibrinolytic agent for the treatment of cardiovascular disease.

Siderophores: Enhancing crop nutrition and improving stress tolerance through microbial iron scavenging 

Parul Sharma, Geeta Rawat, Prerna Gupta, Bindu Devi, Diksha Nautiyal

APJMBB 33(2): 183-196

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.18

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Soil stress conditions, such as nutrient deficiencies, drought, salinity, and heavy metal contamination, significantly hinder plant growth and agricultural productivity. This review explores the use of microbial consortia, particularly siderophore-producing bacteria, as biocontrol agents and biofertilizers to mitigate these stresses. Siderophores, molecules that sequester iron, enhance plant access to nutrients while limiting pathogen growth. The microbial consortia improve nutrient availability, combat pathogens, and stimulate plant growth. These microorganisms also enhance soil fertility, resistance to abiotic stress, and overall soil health. The review examines the molecular mechanisms behind siderophore production, their chemical structures, and the interaction between plant roots and microbes. It also highlights the potential of siderophores to promote plant growth on degraded lands, offering an alternative to chemical fertilizers. This approach represents a shift towards sustainable, biologically-driven agricultural practices, reducing reliance on synthetic inputs. However, the exact mechanisms of siderophore action remain unclear. The review identifies research gaps in understanding their production, modes of action, and roles in soil-plant-microbe interactions. Addressing these gaps could lead to optimized siderophore-based consortia, providing sustainable solutions for mitigating soil stress, improving soil health, and boosting agricultural productivity.

Carpaine modulates calcium mobilization through PLC-InsP3 pathway in cardiomyocyte 

Nur Syafinaz Wasli, Eleanor Joanne Muyou, Suhaini Sudi, Siat-Yee Fong, Caroline Sunggip

APJMBB 33(2): 197-204

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.19

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Carpaine, an alkaloid found in the leaves of Carica papaya (papaya), is known for its diverse pharmacological properties. A previous study demonstrated that carpaine promotes cell proliferation and repair of H9c2 cardiomyocytes after oxidative insults. However, the exact mechanism of action remains elusive. Calcium (Ca²⁺) is the most versatile and universal signalling agent modulating essential physiological functions. This study aims to decipher the calcium modulation of carpaine extracted from C. papaya leaves. Carpaine was isolated from C. papaya leaves, and nuclear magnetic resonance (NMR) spectroscopy confirmed the extracted alkaloid's purity. The 50% inhibitory concentration (IC₅₀) of carpaine in cardiomyocytes was determined using an MTT assay. Fluorescence-based assays were used to determine intracellular Ca²⁺ mobilization. Pyrazole-3-carboxylic Acid (Pyr3), U71322, and Xestospongin C (Xest C), which are the inhibitors of TRPC3/6, Phospholipase C and the InsP3 receptor, respectively, were used to explore the mode of calcium mobilization in cardiomyocytes. Carpaine induced a significant increase in Ca²⁺ mobilization into cardiomyocyte cells. Inhibition of PLC or TRPC3 significantly suppresses the Ca²⁺ mobilization, but interestingly, the inhibition of the InsP3 receptor alone nearly abolished the carpaine-induced Ca²⁺ mobilization. These findings suggest that carpaine induces a calcium store-dependent intracellular Ca²⁺ mobilization in cardiomyocytes. 

Effects of different levels of cave bats guano on chilli plant growth

Siti-Syamim Nurfatihah Binti Abd Rahman, Mohamad Fhaizal Mohamad Bukhori

APJMBB 33(2): 205-212

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.20

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Chilli plants (Capsicum annuum) from the Solanaceae family are among the most widely cultivated crops globally. However, chilli plants are susceptible to numerous threat that can limit production, including pests, diseases, and environmental stressors that cause flower and fruit drop. Fertilization is critical for optimizing chilli yields, with studies showing that the application of fertilisers can substantially improve plant growth and yield. Bat guano, a highly nutrient-dense organic fertiliser rich in nitrogen, phosphorus, potassium, carbon, and beneficial microbes, has been used for centuries to improve crop growth in various countries. The present study investigates the effects of cave bat guano on several growth parameters in chilli plants, including plant height, leaf count, flower production, and fruit yield. For this study, guano was collected from Wind Cave Nature Reserve and Raya Cave in Sarawak. The experiment involved treating 81 chilli plants with three different guano levels (2.5 g, 5.0 g, and 7.5 g), along with a control group that received no treatment. Plant height, leaf count, and counts of flower buds, flowers, and fruits were measured monthly. Bat guano, especially at a rate of 7.5g per plant, was found to substantially improve chilli plant growth, flowering, and fruiting. The control group, by contrast, exhibited stunted growth, underscoring the importance of nutrient supplementation in achieving optimal growth. These findings are in line with prior research on guano applications, establishing it as a valuable organic fertiliser with the potential to boost agricultural productivity in Malaysia.

Impact of high fat diet on the kidney proteome of diabetic rats: A 2D-gel electrophoresis and LC-MS/MS study

Xuan-Yi Sim, Lay-Harn Gam

APJMBB 33(2): 213-223

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.21

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Diabetes can lead to kidney diseases, in this study we evaluated the impact on diabetes under the influence of high fat diet on the kidney by analysing the changes in kidney proteome, this is because proteins play a critical role in kidney function.  Kidney tissue proteomics was carried out by means of 2-dimensional gel electrophoresis LC-MS/MS analysis. The diabetic rats were divided into 4 groups (n=6), namely, G1 (diabetic fed with normal diet), G2 (diabetic fed with normal diet and metformin treatment), G3 (diabetic fed with high fat diet), G4 (diabetic fed with high fat diet and metformin treatment). The proteins that were expressed in ≥ 2-folds (p< 0.05) in pairwise comparison were alpha-enolase, heat shock protein 90, and prohibitin in diabetic rats fed with high fat diet (G3 vs G1); beta subunit haemoglobin in diabetic rats fed with normal diet and treated with metformin (G2 vs G1); haptoglobin in diabetic rats fed with high fat diet and treated with metformin (G4 vs G3).  Significant (p<0.05) upregulation of alpha-enolase, HSP90 and prohibitin while downregulation of carboxypeptidase in the kidney tissue of diabetic rats fed with HFD concurrently pointing to the inflammation of the rats’ kidney.  Elevation of haemoglobin and haptoglobin levels in metformin treated rats was functioned to amend kidney injury.

Advancement in therapeutic approach for treatment of metastatic colorectal cancer

Singireddy Shreya, Raghavendra B N, Sudhakar Kancharla, Prachetha Kolli, Gowtham Mandadapu,
Manoj Kumar Jena

APJMBB 33(2): 224-238

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.22

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Metastatic colorectal cancer (m-CRC) is an advanced cancer type that occurs when cancer cells travel from the colon or rectum to other organs like lungs or liver. The cause of m-CRC is due to multifactorial variables which include genetic anomalies, tumour heterogeneity and factors within the tumour microenvironment (TME). The m-CRC poses a substantial clinical challenge that calls for the development of novel therapeutic approaches. This review discusses in details about the most recent developments in targeted and personalised therapeutic strategies for m-CRC. The necessity of tailored treatment plans based on patient-specific characteristics and molecular profiles, highlights the importance of biomarkers in maximising therapeutic success. The review also discusses the difficulties in treating the m-CRC, such as overcoming resistance to targeted medicines. Recent researches and historic trials have provided new dimensions on the therapeutic approaches towards m-CRC, emphasising the urgent need for customised therapies to improve patient survival and responsiveness towards therapy. The need for considering liquid biopsies and circulating tumour DNA (ctDNA) in treatment decisions is also discussed. Furthermore, other novel therapies such as the combination of VEGF inhibitors with chemotherapy for enhancing treatment efficacy, CAR-T cell therapy, and bispecific antibodies are also considered.

Erratum - Evaluation of the cytotoxic activity of essential oils from Malaysian herbal plants against A375 and A431 skin cancer cell lines

Bassam Mohamed Jawahar, Raha Ahmad Raus, Munira Shahbuddin, Mohd Rushdi Abu Bakar, Nur Najieha Binti Mohd Pauzi, Dayang Fredalina Basri, Irna Elina Ridzwan

APJMBB 33(2): 239

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2.23

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The corresponding author recognized an error in the funding grant code and requested a correction from FRGS21-217-0826 to FRGS/1/2021/STG05/UIAM/02/3. This amendment is necessary to ensure that the published article is recognized as an output of the grant. 

Volume 32 (2) Supplementary Issue

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This supplementary issue contains abstracts presented at International Conference on Molecular Biology and Biotechnology 2025 (ICMBB2025)

Volume 33(2) (Special Issue); 2025

Therapeutic potential of Tetracera scandens on oxidative stress using aspartame-induced oxidative stress rat model and toxicity assessment

Amyra Amat Sain, Mohd Hasnan Mohd Noor, Md Azman Seeni Mohamed, Zafarina Zainuddin

APJMBB 33(2) (Special Issue): 1-15

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2(Special).01

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Tetracera scandens Linn. Merr., a medicinal plant from the Dilleniaceae family, is native to Southeast Asia and has been traditionally used to treat various ailments, including diabetes and inflammation. This study aimed to evaluate the therapeutic effects and subchronic toxicity of an aqueous extract of T. scandens leaves (TSLAE) on oxidative stress (OS) in male Wistar albino rats. The hypothesis posited that TSLAE would mitigate oxidative stress-related damage. Male rats were divided into four groups: a control group (reverse osmosis water), an aspartame (ASP) control group, and two treatment groups receiving low (350 mg/kg) and high (700 mg/kg) doses of TSLAE after ASP exposure for 28 days. Key biomarkers, including superoxide dismutase (SOD) levels and lipid peroxidation, were assessed post-treatment. Results indicated that the ASP-induced OS model was successfully established, with no observed mortality during the study. Histopathological analysis revealed no toxic effects on kidney, liver, or brain tissues, and blood biochemistry showed no significant differences among groups, indicating safety. Notably, high-dose TSLAE treatment significantly reduced SOD levels and lipid peroxidation compared to controls, suggesting a restoration of oxidative balance beyond baseline conditions. In conclusion, TSLAE demonstrated a non-toxic profile while effectively ameliorating oxidative stress biomarkers, highlighting its potential therapeutic role in addressing oxidative stress-related neurodegenerative disorders.

Screening of phytochemicals extracted from Leucaena leucocephala by using microwave-assisted hydrodistillation (MAHD)

Nur Farahain Mohd Nasruddin, Siti Nur Nadzmiah Mohd Nor, Wan Norlinda Roshana Mohd Nawi, Nor Hanimah Hamidi

APJMBB 33(2) (Special Issue): 16-22

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2(Special).02

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Leucaena leucocephala is known as ‘Petai Belalang’ in Malaysia and can be identified by its intermediate leaflets and large pods. The leaflets can be up to 16 mm long and 4.5 mm wide, while the pods occur in a crowded cluster of 5-20 pods per flower head and with 8-18 seeds per pod. Much literature has been published on the medical benefits of L. leucocephala, which exhibits antidiabetic and anti-inflammatory activities. Phytochemical compounds have received considerable attention because of their unique and vast spectrum of biological activities. The extraction of phytochemicals from L. leucocephala was carried out using microwave-assisted hydrodistillation (MAHD) at varying temperatures ranging from 30 °C to 70 °C at 10 °C intervals. The compounds extracted were then identified using gas chromatography-mass spectroscopy (GC-MS). Several bioactive compounds, including glycosides, phenols, terpenoids, and alkaloids, were identified, with glycosides predominantly present in the pods and seeds of L. leucocephala. The highest concentration of these compounds was identified at an extraction temperature of 50 °C, as determined by the peak area in the GC-MS chromatogram. The findings revealed that glycosides were the most abundant phytochemicals present in L. leucocephala, while other compounds, including phenolic acids, steroids, and terpenoids, were identified in lower amounts. In addition, most of the identified compounds exhibited antidiabetic, anti-inflammatory, antioxidant, antibacterial, and antifungal properties, while some compounds also demonstrated therapeutic potential against cancer. Among these compounds, 4-C-methyl- myo-inositol was present in significant quantities and potentially demonstrated both antidiabetic and anti-inflammatory agents. These results indicate that L. leucocephala holds significant promise as a medicinal plant due to its diverse pharmacological activities. 

HPLC quantification of bioactives and antioxidant activity in whey kefir-enriched ecoenzyme from rockmelon (Cucumis melo) and orange (Citrus sinensis) peels

Amyra Hazwani Ghazali, Siti Alwani Ariffin, Wan Zuraida Wan Mohd Zain, Neneng Windayani and Fatimah Salim

APJMBB 33(2) (Special Issue): 23-31

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2(Special).03

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Rockmelon (Cucumis melo) and orange (Citrus sinensis) are rich sources of β-carotene, vitamin C, and polyphenols. In Malaysia, rockmelon production reached 20,000 metric tons (1,492 hectares, 2021), while orange production increased by 52.8%, reaching 14,900 metric tons in 2022. Their peels, comprising 30–35% and 50–65% of the fruit weight, respectively, are significant agricultural wastes containing  phenolic acids, flavonoids, alkaloids, and carotenoids. Whey, a dairy byproduct making up 80–90% of milk volume from cheese and kefir production, is often discarded, even though it contains lactic acid, bioactive peptides, and probiotics, which offer antioxidant potential for sustainable agriculture. This study aimed to analyze the phytochemical compounds and assess the antioxidant activity in rockmelon and orange peel ecoenzymes enriched with whey kefir through a three-month fermentation process using high-performance liquid chromatography (HPLC), radical scavenging assays (DPPH), total antioxidant capacity assays (FRAP), total phenolic content (TPC), and total flavonoid content (TFC). HPLC analysis identified ascorbic acid, gallic acid, tannic acid, and p-coumaric acid were detected. These bioactive compounds significantly enhanced the antioxidant capacity of the EROW50% ecoenzyme, achieving an IC50 of 0.57 mg/mL in DPPH assays and TPC values of 9.03 mg GAE/g. These findings indicate that whey kefir effectively enhances the antioxidant potential and bioactive content of ecoenzymes from fruit peels. The study concludes that whey kefir is a valuable additive for improving the efficacy of natural antioxidants in ecoenzyme formulations, highlighting its potential for developing potent, sustainable products from agricultural waste.

Phytochemical composition, antioxidant potential, and antimicrobial applications of ecoenzymes: A comprehensive review

Wan Zuraida Wan Mohd Zain, Amyra Hazwani Ghazali, Siti Alwani Ariffin, Afiqah Mohammad, Nur Sakinah Asya Ismail and Neneng Windayani

APJMBB 33(2) (Special Issue): 32-41

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2(Special).04

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The increasing resistance to synthetic antimicrobial and the need for sustainable waste management solutions have led to the exploration of ecoenzymes as an alternative. Ecoenzymes, produced through the fermentation of organic waste such as vegetable and fruit peels, have gained attention for their dual role as antimicrobial agents and waste management tools. This review critically examines the phytochemical composition, antioxidant activity, and antimicrobial properties of ecoenzymes, positioning them as a novel, sustainable alternative to conventional antimicrobial and chemical treatments. The data was collected from international and national indexed journals and proceedings through a systematic literature review using Scopus, PubMed and Google Scholar with advanced search terms such as "ecoenzyme," "antimicrobial activity," "antioxidant activity," and "phytochemical profile." Studies utilising ecoenzymes from various organic materials, including Cucumis melo, Citrus sinensis, Carica papaya L., Azadirachta indica, Ananas comosus, etc were analysed. The findings demonstrate that ecoenzymes are rich in bioactive compounds such as flavonoids, tannins, and saponins, which contribute to their antioxidant capacity. Antimicrobial testing showed that ecoenzymes effectively inhibit a range of pathogenic bacteria, primarily through mechanisms that disrupt bacterial cell membranes and inhibit nucleic acid synthesis. Additionally, ecoenzymes are found to have potential in reducing the environmental impact by transforming organic waste into valuable antimicrobial agents. Ecoenzymes offer a promising, sustainable alternative to synthetic antimicrobials while simultaneously addressing waste management challenges. The review highlights the potential for ecoenzymes to be integrated into applications such as healthcare, agriculture, industry and environmental management, promoting the circular economy and reducing reliance on harmful chemicals.

Isolation, structure elucidation, and antibiofilm activity of α-viniferin from Hopea ferrea bark extract

Mohd Fithriee Zailani, Nurul ‘Izzah Mohd Sarmin, Isna Athirah Othman and Fatin Nadiah Thaqifah Shahriman

APJMBB 33(2) (Special Issue): 42-47

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2(Special).05

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Hopea ferrea, a species within the Dipterocarpaceae family, has long been recognized for its traditional medicinal applications. The aim of this study is to isolate and elucidate the structure of α-viniferin from the bark of H. ferrea and to evaluate its antibiofilm activity against Streptococcus mutans.  The isolation process was conducted using Medium Pressure Liquid Chromatography (MPLC) and High-Performance Liquid Chromatography (HPLC), resulting in the successful purification of α-viniferin. Structural elucidation was confirmed in Nuclear Magnetic Resonance (NMR) and comparison with literature data. The biofilm inhibitory potential of α-viniferin was evaluated against S. mutans, a major contributor to dental caries. Our findings demonstrated that α-viniferin inhibits biofilm formation in S. mutans, highlighting its potential for development as a natural agent in oral healthcare applications.

Renal and hepatic toxicological impact of orally administered Dioscorea hispida aqueous extract in rats 

Nik Aina Syazana Nik Zainuddin, Nur Liana Md Nasir, Maizatul Hasyima Omar, Hussin Muhammad,
Norizah Awang

APJMBB 33(2) (Special Issue): 48-53

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.2(Special).06

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Despite being an under-reported toxicity, Dioscorea hispida (D. hispida) or ubi gadung is a tropical yam that is commonly used as a traditional remedy and a source of carbohydrates in Malaysia. Various studies have reported its toxicological effects. However, the impact of the administration of this yam towards kidneys and liver has not yet been widely reported. Hence, this study was conducted to investigate the toxicological effect of D. hispida aqueous extract on the kidney and liver of Sprague Dawley rats through histopathological examination. Forty Sprague Dawley rats (n = 20 males and n = 20 females) were randomly divided into four groups consisting of a control group (distilled water) and three D. hispida aqueous extract groups (125 mg/kg, 500 mg/kg and 2000 mg/kg BW). The extract was orally administered daily for 28 days, and all rats were sacrificed at the end of the study. The livers, left and right kidneys were collected for histopathological examination via hematoxylin and eosin (H&E) analysis and the lesions observed were scored accordingly. All collected organs showed moderate congestion and necrosis while mild severity for haemorrhage, degeneration and inflammation in all treated groups compared to the control group. The necrosis of both kidneys of rats was seen on the glomerular, medullary and cortex regions with loss of normal architecture in kidneys for the 2000 mg/kg BW group. Significant changes (p<0.05) were observed in the scoring of necrosis on both kidneys of rats treated with 500 mg/kg and 2000 mg/kg BW compared to the control group. Apart from necrosis and congestion, infiltration of inflammatory cells was also observed in the liver. However, scores for observed lesions on the liver of rats were not significant for all treated groups. In conclusion, administration of D. hispida aqueous extract for 28 days exerted renal toxicity at 500 mg/kg and 2000 mg/kg BW, but no toxicity was observed on the liver at all doses of rats.

Volume 33(3); 2025

Salinity and foliar application of methyl jasmonate change the expression of the heat shock protein 90 (Hsp90) gene and morphological traits during the reproductive stage of rice
(Oryza sativa)

Zairul Fazwan Md Zainordin, Cha Thye San, Aziz Ahmad

APJMBB 33(3): 1-14

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.01

Click here to download: [PDF] [Supplementary File]

Salinity is one of the problems in rice growth and yield. Studies on the effect of exogenous plant growth regulators on salinity-stressed plants remain limited. Therefore, this study aimed to investigate the impact of salinity stress and methyl jasmonate (MeJA) foliar application on growth characteristics and OsHsp90 gene expression in three rice cultivars (MR219, MR219-4, and MR219-9). Suggesting that the combined application of MeJA and activation of the OsHsp90 gene may have synergistic effects in enhancing salt tolerance. The Real-Time PCR was used to quantify the expression level of the gene in the leaves from the rice plants at the early reproductive stage. The plant growth parameters were measured 100 days after planting. Results showed that the expression of the OsHsp90 gene in the leaves of rice plants to MeJA treatment and salinity conditions varied among the rice varieties. The finding indicates that the MR219-9 variety exhibits greater tolerance and better growth in saline conditions than the other two genotypes (MR219 and MR219-4). Notably, a significant up-regulation expression (p < 0.05) of the OsHsp90 gene levels in the MR219-9 genotypes under the MeJA + salinity treatment (21.41 ± 5.34-fold) suggests its potential role in responding to stress conditions. This study provides valuable insights into the role of exogenous plant regulators, particularly MeJA, in mitigating the effects of salinity stress on rice plants. Future research should investigate the impact of other plant growth regulators on the expression of various genes in rice plants grown under saline conditions to improve the growth and production.

The association of screen exposure, melatonin suppression and OPN4 gene polymorphism in children with speech delay: A review

Donna Hermawati, Agustini Utari, Tri Indah Winarni

APJMBB 33(3): 15-23

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.02

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Speech delay is a developmental condition in which children experience delayed speech and language acquisition without concurrent impairments in other domains. The prevalence of speech and language delay varies from 2.3% to 19% worldwide. Speech delay is caused by various risk factors, such as environmental, perinatal, and genetic factors. Excessive exposure to electronic media is one of the important environmental components connected to hyperactivity, attention deficits, and delayed language development. Blue light exposure from screens affects melatonin, a crucial neuroplasticity and sleep regulator that may affect language acquisition and brain connectivity. In addition, the Opsin4 (OPN4) gene is responsible for encoding melanopsin and plays a crucial role in mediating non-image-forming visual tasks and regulating physiological processes in response to light. Polymorphisms in the OPN4 gene may alter melanopsin signalling pathways, influencing circadian rhythms and melatonin regulation. OPN4 genetic variants such as the P10L and I394T polymorphisms may impact melanopsin and neurophysiological functions and sleep patterns. Excessive screen exposure further disrupts melanopsin-driven responses, contributing to sleep disturbances and decreased melatonin levels, which may indirectly impact speech and cognitive development. Further research is essential to a better understanding of complex interactions between screen exposure, genetic predisposition, and environmental influences on speech delay. Raising awareness about these impacts can help caregivers and educators promote balanced screen use and create healthier developmental environments, especially for young children.

Proof-of-concept of duplex MNAzyme assay for simultaneous detection of Candida auris and Candida spp. using synthetic
DNA targets

Mohd Hanif Jainlabdin, Muhanna Mohammed Ahmed Al-Shaibani, Mosaad Attia Abdel Wahhab, Mohammad Ridhuan Mohd Ali, Thomas Edwards, Jorge Tovar-Torres

APJMBB 33(3): 24-30

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.03

Click here to download [PDF] [Supplementary File]

The global emergence of Candida auris as a multidrug-resistant fungal pathogen presents significant challenges to healthcare systems, particularly in hospital settings. Rapid and accurate diagnostics are crucial for managing C. auris infections. This study presents a preliminary analytical evaluation of a duplex Multicomponent Nucleic Acid Enzyme (MNAzyme) assay designed to simultaneously detect C. auris and genus-wide Candida species using synthetic DNA targets. Target sequences were selected from the conserved D1/D2 region of the LSU rDNA locus, and melting temperature analysis confirmed distinct probe hybridisation profiles, with Substrate 2:PCO1 yielding a Tm of 45°C and Substrate 3:PCO4 at 26°C. The assay demonstrated specificity by differentiating C. auris from other Candida species through melting curve analysis. In reactions containing synthetic targets, probe–target binding disrupted probe–PCO hybrids, resulting in a marked reduction in melt peaks compared to no DNA controls (NDC). The duplex format allows multiplex detection within a single fluorescence channel, offering a potential advantage over conventional single-target assays. This proof-of-concept establishes a rapid, low-cost diagnostic platform with promising potential for future clinical application. Further validation using clinical and environmental samples is needed to support its broader utility in detecting fungal pathogens.

Identification of virulence genes and antimicrobial susceptibility of Escherichia coli: comparison between plaque psoriasis and healthy donor faecal samples

Nur Insyirah Mohd Razalan, Shanthi Krishnasamy, Barathan Muttiah, Mohan Arumugam, Navindra Kumari Palanisamy, Kumutha Malar Vellasamy, Vanitha Mariappan

APJMBB 33(3): 31-39

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.04

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Psoriasis is associated with gut microbiota dysbiosis, which studies link with increased Escherichia coli in the gut. Coherently, E. coli also develops resistance to commonly prescribed antibiotics. This study aimed to isolate, identify, and confirm E. coli isolated from faecal samples of plaque psoriasis (mild and severe) patients and healthy donors that match the patients' in terms of age range, gender, and race. MacConkey agar was used to isolate the E. coli from faecal samples, and DNA was extracted using the boiling method. Genotypic confirmation of E. coli using the 16S rRNA gene was performed before identifying the presence of virulence genotyping. Antimicrobial susceptibility testing (AST) was conducted to determine the presence of multidrug-resistant (MDR) E. coli. Virulence genotyping analysis revealed that only the toxin gene (sat), iron acquisition gene (fyuA) and the capsule synthesis gene (kpsmTII) were found in E. coli isolated from psoriasis patients compared to healthy donors. All 20 E. coli isolates were susceptible to gentamicin (10 µg) and ceftriaxone (30 µg), but either resistant or intermediate to imipenem (10 µg), ampicillin (10 µg), and erythromycin (15 µg). Four out of 20 tested isolates were MDR E. coli. These findings suggest that virulence genes may have played a role in the development of psoriasis. Treatments for E. coli infections point to the need for prior AST to combat antibiotic resistance while effectively navigating patients' treatment courses.

The effect of Phyllanthus debilis methanolic extract on miR-125a-5p and miR-320a expression in HT-29 colorectal cancer cell line

Nurdianah Harif Fadzilah, Siti Nur Dalila Mohd Zain, Wan Adnan Wan Omar

APJMBB 33(3): 40-45

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.05

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miR-125a-5p and miR-320a expressions were often dysregulated in colorectal cancer. Using HT-29 cell line as a model of colorectal cancer, we hypothesized that Phyllanthus debilis (P. debilis) methanolic extract could regulate the expression of miR-125a-5p and miR-320a, thereby altering the progression of colorectal cancer. The cell culture study on the effect of P. debilis methanolic extract on miRNA expression was conducted using the HT-29 cell line. The effect of P. debilis methanolic extract on the proliferation of HT-29 cells was dose-dependent, with a half-maximal inhibitory concentration (IC₅₀) of 0.1 mg/mL. Cells were treated with 0.1 mg/mL P. debilis methanolic extract and 0.5 µM 5-Aza-2-Deoxycytidine (positive control) for 72 h. Cells were then harvested at 72 h, and RNA was extracted. The expressions of miR-125a-5p and miR-320a were measured using TaqMan® assays and quantified using Reverse Transcription-Quantitative Real-Time PCR (RT-qPCR). The results showed that the treatment with P. debilis methanolic extract significantly increased the expression of miR-125a-5p by 2.3-fold and miR-320a by 1.6-fold (p < 0.05) compared to untreated HT-29 cells. In contrast, treatment with 5-Aza-2-Deoxycytidine resulted in the opposite effect, significantly decreasing the expression of miR-125a-5p by 0.5-fold and miR-320a by 0.5-fold (p < 0.05). The anticancer effect of P. debilis may be partly regulated through miRNA regulation. 

Expression, purification, and substrate specificity of recombinant zebrafish D-amino acid oxidase 

Ming-Kai Chern, Yun-Hsin Wang, Mei-Yun Huang, I-Ching Kuan, Bo-Chang Wang, Yau-Hung Chen

APJMBB 33(3): 46-50

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.06

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D-amino acid oxidase (DAO) is a peroxisomal enzyme that catalyzes the oxidative deamination of D-amino acids. In this study, we employed a bacterial expression system to express and purify a ~38 kDa recombinant zebrafish DAO (zDAO) protein. The kinetic parameters, including Km (μM), Kcat (nmol/min/mg), and substrate specificity (Kcat/Km), were determined. All D-amino acids were tested as substrates. Among them, zDAO exhibited the highest specificity for D-alanine (0.537), making it the preferred substrate. In contrast, D-threonine showed the lowest specificity (0.018), approximately 3% of that for D-alanine. These findings provide further insight into the enzymatic properties of fish DAO. 

Application of CRISPR-Cas9 technology in farm animals:
A comprehensive review

Palivela Pranay Pritham, Tadikonda Sai Siri, Geethika Madala, A. Hema Sindhuri, Sudhakar Kancharla, Prachetha Kolli, Gowtham Mandadapu, Prasana Kumar Rath, Bidyut Prava Mishra, Manoj Kumar Jena

APJMBB 33(3): 51-65

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.07

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Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) - Cas9 technology has transformed genetic engineering, particularly in the domain of farm animal production, offering precise and efficient genome editing capabilities. This comprehensive review article discusses in details about the diverse CRISPR-Cas9 applications in livestock, highlighting its potential to enhance disease resistance, improve reproductive health, and increase productivity. Major advancements include the development of pigs resistant to Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) by targeted modifications of the CD163 gene, which reduces the need for antibiotics and vaccines. In poultry, CRISPR has facilitated the creation of chickens resistant to avian influenza, demonstrating its effectiveness in controlling infectious diseases. Additionally, technology permits the introduction of desirable traits, like hornless characteristics in cattle, thereby eliminating the necessity for painful dehorning procedures. Despite these promising applications, the utilization of CRISPR-Cas9 brings forth significant ethical issues regarding biodiversity, animal welfare, and the lasting impacts of genetic alterations. Technical challenges, encompassing the possibility of unintended effects and the need for efficient delivery mechanisms, must also be addressed to ensure the safe application of this technology. As research in genome editing for farm animals progresses, it is essential to balance scientific innovation with ethical considerations and regulatory frameworks. This review insights the importance of ongoing studies to optimize CRISPR-Cas technology. Effective integration of CRISPR-Cas9 in livestock breeding significantly enhances animal health status, food security, and contributes to sustainable agricultural production, addressing the increasing global demand for food while minimizing environmental impacts. The review article with updated information, will help the researchers working in this area to design their future research work efficiently. 

Genetic diversity and population structure of Azolla inferred by trnL-trnF intergenic region and morphological data in Yogyakarta, Indonesia

Aji Sukma Iqbal Najibulloh, Adib Fakhruddin Yusuf, Abdul Razaq Chasani, Budi Setiadi Daryono

APJMBB 33(3): 66-77

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.08

Click here to download [PDF] [Supplementary File]

Azolla, commonly known as fairy moss, is a floating aquatic fern found in both tropical and temperate regions. While it plays an important ecological and agricultural role—such as enhancing soil nutrients in rice paddies—it has also been identified as an invasive species in many freshwater ecosystems. Its spread is often associated with the aquarium plant trade and the interconnectedness of freshwater systems. Understanding the genetic variation and population structure of Azolla is essential for monitoring its spread and managing its ecological impact. This study aimed to analyze the distribution and population structure of Azolla species collected from both wild (rice fields) and cultivated (ponds) habitats in the Special Region of Yogyakarta, Indonesia. Morphological observations were conducted alongside molecular analyses using DNA sequences from the intergenic spacer region trnL-trnF. Phylogenetic analysis revealed that Azolla populations in Yogyakarta are grouped into two distinct clades, with genetic divergence values ranging from 0.00% to 0.74%. Each clade exhibited a unique haplotype with low nucleotide diversity. All samples were identified as Azolla cristata (conspecific with A. microphylla and A. mexicana) with 90% confidence. Clade groupings corresponded to habitat type, distinguishing wild from cultivated populations. Haplotype network analysis showed overlapping distribution patterns, likely influenced by the connectivity of regional freshwater systems. These findings contribute to the understanding of Azolla diversity in Indonesia and provide a foundation for future monitoring of its genetic structure. Broader population sampling is necessary to further evaluate potential invasiveness or the risk of biodiversity loss in freshwater environments.

In vitro antiviral activity of Coriandrum sativum L. seed crude extracts against respiratory syncytial virus

Darssheela Ramasamy, Nur Suhanawati Ashaari, Zetty Norhana Balia Yusof, Azimah Abdul Wahab, Iffah Izzati Zakaria, Saila Ismail

APJMBB 33(3): 78-94

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.09

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Human respiratory syncytial virus (RSV) causes severe lower respiratory infections in young children and the elderly. Currently, no specific antiviral drug exists for RSV. Ribavirin, a broad-spectrum nucleoside analogue originally used for hepatitis C virus, is reserved for severe RSV cases because of its side effects and high cost. This underscores the need for safer, more affordable alternatives. Coriandrum sativum L. (coriander) has shown antiviral activity against several viruses; however, its potential against RSV remains unexplored. This study investigates the antiviral effects of coriander seed extracts against RSV in vitro. Methanolic (MSECS), hexane (HSECS), and aqueous (ASECS) extracts of coriander seeds were tested on RSV-infected BEAS-2B human bronchial epithelial cells. Intracellular viral loads were quantified using end-point PCR, and extracellular infectious titres were assessed via 50% tissue culture infectious dose (TCID₅₀) assay. Gas chromatography-mass spectrometry (GC-MS) was used to identify bioactive compounds in the extracts. At non-toxic concentrations of 62.5 and 125 µg/mL, MSECS, HSECS, and ASECS significantly reduced RSV intracellular RSV loads, comparable to ribavirin control. At 125 µg/mL, MSECS, HSECS, and ASECS decreased extracellular viral titres by 97%, 84%, and 98%, respectively, surpassing ribavirin’s inhibitory effect. GC-MS analysis revealed hexadecanoic acid, octadecanoic acid, linalool, and myristic acid as common major compounds across all extracts. Additionally, MSECS contained geranyl acetate (7.57%), HSECS included capric acid (25.1%), and ASECS was rich in cis-vaccenic acid (60.7%). Some of these compounds are known for their antiviral properties, suggesting a potential synergistic mechanism in RSV inhibition. Our results provide the first evidence that Coriandrum sativum L. crude extracts possess potent in vitro antiviral activity against RSV and may serve as novel, safe, and affordable antiviral agents against RSV.

Tight junction modulation by Bifidobacterium bifidum: A review on the potential mechanisms of lapatinib-induced diarrhoea

Nur Syafina Hisham, Wan Nor I’zzah Wan Mohamad Zain, Jesmine Khan, Narimah Abdul Hamid Hasani, Hassanain Al-Talib

APJMBB 33(3): 95-108

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.10

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Lapatinib is a targeted therapy for ErbB2-positive breast cancer. Despite its efficacy, a notable side effect associated with lapatinib is diarrhoea. Due to the prolonged duration of treatment, diarrhoea significantly impacts patients’ quality of life. A reduction in Bifidobacterium spp. levels in diarrhoea patients treated with tyrosine kinase inhibitors (TKIs) indicates an altered gut bacteria composition. However, the precise mechanisms remain unclear. This review investigates potential mechanisms behind lapatinib-induced diarrhoea through its effect on Bifidobacterium bifidum (BB) in altering the tight junctions (TJs) of the gastrointestinal (GI) tract. Scopus and PubMed searches were performed using current data from 2018 to 2024, with BB, lapatinib, diarrhoea, intestinal permeability, and TJ proteins as keywords. A total of 16 of 167 Scopus articles and 29 of 58 PubMed articles were selectively discussed. The effects of bacterial exposure and alteration of TJs in the GI tract were investigated using the in vitro model Caco-2, as these cells can spontaneously differentiate into polarised monolayers. Overall, these articles highlighted the significance of Bifidobacterium spp. studies, in maintaining intestinal homeostasis. Notably, BB plays an essential role in modulating intestinal flora and enhancing barrier function. Therefore, alterations in BB induced by lapatinib may influence TJ properties, enhance intestinal permeability, and potentially lead to diarrhoea. This review highlights the importance of understanding how lapatinib induced changes to gut microbiota and intestinal barrier function contribute to diarrhoea. Elucidating these mechanisms may help clinicians better understand the pathophysiology of TKI-induced diarrhoea and improve its management.

Molecular identification of photosynthetic bacteria associated with hardcoral Acropora spp. from different depths in Gosong Cemara Karimunjawa Waters using 16S rRNA gene sequencing

Diah Ayuningrum, Maliha Wiaam Rohadatul’aisy, Aninditia Sabdaningsih, Norma Afiati, Pujiono Wahyu Purnomo

APJMBB 33(3): 109-119

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.11

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Acropora spp. corals dominate the waters of Gosong Cemara, Karimunjawa. These corals can live at various depths through the association of photosynthetic bacteria. This research aims to determine the abundance and types of photosynthetic bacteria associated with Acropora spp. corals at different depths.  The method used in this research is the explorative descriptive method. This research was conducted from November 2023 to March 2024.The results of this research showed that the highest bacterial abundance in medium R2A (1/5 strength) was found in Acropora coral at a depth of 1 meter, with 7.4 × 10¹ CFU/ml. At a depth of 5 meters, the abundance was 6.6 × 10¹ CFU/ml, while at 15 meters, the bacterial count was classified as too few to count (TFTC). Three isolates from R2A medium (1/5 strength) were selected for molecular analysis based on colony color, which indicated potential photosynthetic activity. Molecular identification using the 16S rRNA gene revealed that two of the isolates were photosynthetic bacteria, as indicated by the presence of carotenoid pigments involved in the photosynthesis process. Isolate KJA1-1-2 was identified as Kocuria rhizophila with 99.72% similarity, and isolate KJA2-1-3 as Sphingopyxis terrae with 98.39% similarity.

Metabolomics as a frontier in oral squamous cell carcinoma diagnosis: Understanding metabolic alterations and clinical implications

Pralaya Kumar Sahoo, Sangeeta Chhotaray, Soumya Jal

APJMBB 33(3): 120-131

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.12

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Oral squamous cell carcinoma (OSCC) is a common oral cavity cancer with a high death rate, owing to late detection and limitations in existing diagnostic techniques. Despite advancements in physical examination, imaging, and histopathological techniques, early detection of OSCC remains challenging due to the asymptomatic nature of early lesions and the invasive nature of traditional biopsies. Metabolomics presents a promising frontier in disease diagnosis by analyzing the dynamic and diverse metabolome of the human body. It offers insights into metabolic alterations associated with OSCC and provides potential biomarkers for early detection, prognosis, and monitoring of treatment efficacy. This review summarizes the current state of metabolomics research in OSCC, highlighting significant metabolic changes observed in tumorigenesis, such as altered glucose and amino acid metabolism, and lipid dysregulation. Specifically, this review highlights key metabolic alterations in OSCC, including glycolysis-related metabolites such as lactate and glucose transporters (GLUT1, GLUT3), amino acid metabolism components like glutamine and asparagine, and lipid metabolism markers such as Fatty Acid Synthase (FASN) and cholesterol derivatives. All these metabolites can serve as potential prognostic markers for OSCC. The review concludes with recommendations for future research, including standardizing sample collection methods, focusing on specific metabolites for OSCC, and developing affordable, sensitive diagnostic tools.

Natural products as an alternative therapeutic towards increasing fish survival: a review

Iftikhar Ahmad Abdul Rafi

APJMBB 33(3): 132-142

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.13

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Natural products, which are secondary metabolites produced by plants, have long been used in the treatment of human diseases, with their applications well-documented throughout history. These compounds exhibit a wide range of bioactivities, including anti-inflammatory, anti-parasitic, and anti-bacterial properties. Natural products have been used as alternatives to antibiotics for disease control and treatment aiming to minimize the emergence of bacterial strains with antimicrobial resistance (AMR) which could compromise the effectiveness of future treatments. Furthermore, the usage of antibiotics might expose consumers to health problems, due to their residues in fish products. Besides that, the routine use of chemicals as disinfectants poses long-term health hazards to aquaculture operators, as some of these chemicals are harmful and potentially carcinogenic. Natural products have been shown to effectively treat bacterial and parasitic diseases in aquaculture. Furthermore, the use of natural products in fish feed have also increased their survival against pathogens and improved fish growth. However, since plants typically produce their active components in minute quantities, there is a growing need to produce analogues with similar bioactivities. Chemical synthesis of these active component analogues will enable mass production of these analogues for extensive use in aquaculture industries as commercial products. The future of these naturally derived antimicrobial compounds looks promising, reducing the dependency of antibiotics in aquaculture and ensuring fish products are safe for human consumption.

Protective role of Physalis angulata L. extract in mitigating pulmonary damage from benzo[a]pyrene exposure: A histological study

Nuha Hatem Khalaf, Ghadir Mahmmoud Nejim Al_Rubaie

APJMBB 33(3): 143-149

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.14

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The study explores the defensive properties of Physalis angulata L. (Solanaceae) in preventing lung injury caused by exposure to Benzo[a]pyrene (B[a]P), a carcinogen known to cause significant mutagenic DNA adducts. The focus is on the anti-inflammatory and antioxidant properties of P. angulata, which contain phenols that may alleviate the oxidative stress associated with B[a]P exposure. This study highlights the tissue changes that occur in the lungs of mice exposed to B[a]P with or without application of P. angulata. This process was carried out by continuous histological analysis with the help of a light microscope. The results indicated that there was a significant damaged to the ling tissue such as vascular congestion and destruction of the alveoli, which was caused by B[a]P. Similarly, there was a noticeable improvement in the lung tissue and a reduction in damage, with a normal lung structure being achieved after treatment with P. angulata. This protective effect is due to the active antioxidant and anti-inflammatory properties present in this plant such as physalis and flavonoids, which played a major role in reducing the pneumonia caused by B[a]P. Overall, the study focuses on demonstrating the potential of natural plant extracts to reduce carcinogens from B[a]P environmental toxins, thereby identifying an advanced approach to support overall lung health from widespread toxic exposures.

Development of a novel one-step thermostable heptaplex PCR assay for the detection of medically important Candida species 

Nor Suhada Anuar, Nik Yusnoraini Yusof, Dinie Adila Zainol, Anizah Rahumatullah

APJMBB 33(3): 150-161

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.15

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Invasive fungal infections, particularly candidemia, pose a significant public health challenge due to their high mortality rates. While Candida albicans remains the most common species in candidiasis, there has been a notable rise in non-albicans Candida (NAC) infections, each exhibiting different drug susceptibilities. This underscores the urgent need for rapid, user-friendly detection methods. Conventional molecular tests are limited by the need for expertise, cold-chain logistics, and specific storage conditions, restricting their widespread use. In this study, we developed a thermostabilized multiplex PCR assay to identify five medically significant Candida species: C. albicans, C. krusei, C. glabrata, C. tropicalis, and C. parapsilosis. This assay utilizes newly designed species-specific primers targeting the ITS1 and ITS2 regions. An Internal Amplification Control (IAC) using a specific primer for the SSP2 gene of P. falciparum (273 bp) was included to ensure accuracy. The assay demonstrated 100% sensitivity and specificity when tested with 63 genomic DNA templates from clinical isolates. In addition, the developed thermostabilized heptaplex PCR assay showed stability for at least 6 months as validated by Q10 method testing. In conclusion, this study demonstrated the development of a cold-chain-free PCR assay for the rapid, easy, and simultaneous detection of multiple Candida species. This advancement is intended to improve diagnostic efficiency, support targeted antifungal therapy, and enhance patient outcomes, particularly in resource-limited settings.

Recent advances in biodegradable membranes for oil spill remediation: mechanisms, fabrication, and challenges

Shaheer Alhussain, Larsen Alessandro, Joni Aldilla Fajri, Michelle Oi Yoon Soo

APJMBB 33(3): 162-170

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.16

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Oil spills are highly destructive to marine ecosystems and biodiversity. In addition to the economic loss, there is a dire need for effective, efficient, and sustainable remediation methods to counter the ecological effects of oil spills into open waters. Biodegradable membranes made from cellulose and polylactic acid present a novel alternative to traditional methods. These naturally hydrophobic-oleophilic membranes degrade oil molecules into non-toxic by-products and reduce environmental pollution damage. Fabrication techniques such as electrospinning and phase separation further enhance their efficiency. However, challenges remain in balancing durability with biodegradability and scaling up production. This review highlights the development, challenges, and future perspectives of biodegradable membranes to improve oil spill remediation and promote global environmental sustainability.

Optimization of the SA-β-galactosidase assay for an enhanced senescence detection in H2O2-induced human umbilical vein endothelial cells (HUVECs)

Mas Atikah Lizazman, Vivien Jong Yi Mian, Nur Hisam Zamakshshari, Dayang Erna-Zulaikha

APJMBB 33(3): 171-181

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.17

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Cellular senescence in endothelial cells (EC) plays a critical role in various age-related diseases, including cardiovascular disorders. Senescence-associated β-galactosidase (SA-β-gal) activity, detectable via cytochemical staining with X-gal, remains a widely accepted marker for identifying senescent cells. However, inconsistencies in senescence induction protocols hinder reproducibility and limit their utility in screening anti-senescence compounds. To date, no standardized in vitro conditions have been established for reliably inducing senescence in ECs. Herein, we aimed to utilize H₂O₂ as the primary inducer of cell senescence and investigated the effects of various concentrations, exposure time, chemical grade, and cell culture supplementation in the induction of senescence in human umbilical vein endothelial cells (HUVECs). In this study, we systematically evaluated a range of H₂O₂ concentration, exposure durations, chemical grades, and fetal bovine serum (FBS) supplementation to determine their effects on senescence induction. We quantified senescence by counting SA-β-gal-positive cells under a microscope after X-gal staining. Image-based cell counts, and statistical analyses were conducted to evaluate significance. We observed a threefold rise in SA-β-gal positivity after 24 hours of treatment with 0.5 µM H₂O₂ relative to the control. This indicates that optimal H₂O₂ concentration and exposure duration parameters significantly enhanced senescence. However, neither FBS supplementation nor H₂O₂ chemical grade influenced the outcomes. To conclude, this study highlights the need for an optimized senescence induction method to ensure reliable and interpretable results of SA-β-gal staining in HUVECs. Such improvements will facilitate future investigation of novel pharmacological agents with anti-senescence potential.

Intestinal protozoan infections in Malaysia: Insights into epidemiology, diagnostic approaches, and control strategies:
A mini-review

Shazlina Mizan, Seok Mui Wang and Hassanain Al-Talib

APJMBB 33(3): 182-192

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.18

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Intestinal protozoal infections are a significant public health challenge in Malaysia, especially among vulnerable populations. This article provides a comprehensive literature review on the epidemiology, diagnosis and control strategies of intestinal protozoal infections in Malaysia. The findings highlight the variability in prevalence across different demographic groups and regions, advances in molecular diagnostic methods and ongoing efforts to improve control measures through public health interventions. By identifying research gaps and potential innovations, this review provides valuable insights for policy makers and researchers seeking to curb the impact of intestinal protozoal infections in Malaysia. The prevalence of intestinal protozoa in Malaysia remains remarkably high. This emphasises the urgent need for a range of control and prevention measures. The implementation of effective strategies and the development of multiple diagnostic methods are essential.

Expression and prognostic value of miR-26b-5p in Triple-Negative Breast Cancer (TNBC): Evidence from patient tissues and in silico analysis

Nurhanis Fasihah Muhamad, Md Salzihan Md Salleh, Maya Mazuwin Yahya, Wan Mohd Nazri Wan Zainon, Ahmad Aizat Abdul Aziz

APJMBB 33(3): 193-201

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.19

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Background: Triple-negative breast cancer (TNBC) is associated with a higher likelihood of early tumour relapse and reduced overall survival. MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate gene expression by binding to target mRNAs, leading to transcript degradation or inhibition of translation. This study aimed to evaluate whether miR-26b-5p expression could serve as a prognostic biomarker for predicting recurrence risk and survival outcomes in TNBC. Methods: Total RNA was extracted from tumour and normal adjacent tissues of TNBC patients. The expression level of miR-26b-5p was quantified using RT-qPCR. Potential target genes of miR-26b-5p were identified and survival analysis was performed using METABRIC and TCGA TNBC patient cohorts. Results: The expression level of miR-26b-5p was significantly downregulated in tumour compared to normal adjacent tissues (-4.64 vs -2.29, p=0.001) of TNBC patients. Significant downregulation of miR-26b-5p was also observed in TNBC, which had positive axillary lymph nodes compared to negative axillary lymph node metastases (-2.80 vs -0.0454, p=0.049) as well as in TNBC patients who had disease recurrence compared to non-disease recurrence groups (-3.908 vs -1.536, p=0.009). Survival analysis demonstrated that high miR-26b expression was significantly associated with improved survival rate and high SMAD1 expression was correlated with an increased risk of relapse and poorer survival rates. Conclusion: miR-26b-5p is significantly downregulated in TNBC tumours, particularly in patients with lymph node metastasis and disease recurrence. Its low expression correlates with poorer survival, highlighting its potential as a prognostic biomarker for TNBC recurrence risk.

Metabolomic characterization of different types of Malaysian edible bird’s nest

Rozaihan Mansor, Danish Adli Zulkifli, Aini Ideris, Intan Safinar Ismail, Faridah Abas,
Mokrish Ajat, Nazhan Ilias

APJMBB 33(3): 202-212

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.20

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Edible Bird’s Nest (EBN), produced by Aerodramus fuciphagus and Aerodramus maximus, has been widely consumed for its nutritional and medicinal properties. Despite its increasing global demand, variations in its metabolite composition due to geographical origin and processing methods remain inadequately explored. The current study investigates the metabolite profiles of farmed and commercialized EBN to determine the influence of environmental factors, diet availability, and processing on its biochemical composition. It is hypothesized that EBN from different Malaysian regions exhibits distinct metabolite compositions, potentially impacting its functional properties. Farmed EBN samples were obtained from Perak, Kelantan, Johor, and Sarawak, while commercialized EBN was sourced from a local pharmacy. The metabolite extraction process followed standardized protocols, and proton nuclear magnetic resonance (¹H-NMR) spectroscopy was used for analysis. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to evaluate metabolite variations, while antioxidant properties were assessed using DPPH and nitric oxide scavenging assays. Findings revealed significant differences in metabolite composition between farmed and commercialized EBN. Higher acetone and 4-hydroxyproline levels were detected in commercialized EBN, suggesting potential alterations due to processing. Among farmed samples, Johor’s EBN exhibited the highest nitric oxide scavenging activity, while Kelantan’s EBN demonstrated the strongest DPPH radical inhibition. These variations highlight the impact of geographical and processing factors on EBN’s biochemical properties. Establishing a comprehensive metabolite profile may contribute to industry-wide standardization, ensuring product quality and reinforcing EBN’s potential as a functional food.

Mangosteen (Garcinia mangostana L.) peel extract potential in controlling blood glucose and HbA1c levels in Balb-C mice
by using HPLC

Suharni, Rizki R, Rosmaini 

APJMBB 33(3): 213-221

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.21

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Diabetes mellitus still lacks a perfect therapeutic approach due to the wide variability of its causes and the many complications that occur in people with diabetes mellitus. The management of diabetes mellitus requires a multidisciplinary approach, including pharmacological and non-pharmacological therapies. Mangosteen peel contains Xanthone, which is believed to reduce blood glucose levels or act as an antihyperglycemic. One of the non-pharmacological treatments is the use of mangosteen peel extract. This study investigates the effects of mangosteen peel extract on blood glucose and HbA1c levels in Balb-C mice, aiming to determine whether it can reduce blood glucose and function as an antihyperglycemic agent. The study was designed with a pre-test/post-test to control groups, extracting mangosteen rind and inducing Balb/c mice using alloxan. Each group consisted of 4 mice; the total number of mice to be studied was 25. To account for the dropout, the HbA1c levels were measured using HPLC techniques. There was an effect of mangosteen peel extract on blood glucose levels of Balb-C and HbA1c mice with a significance value of 0.018 <0.05 and a significance value of 0.010 <0.05, respectively. The most effective dose identified in the study is V5 with a dose of 500 mg. 

Investigation of key virulence factors in Candida parapsilosis isolated from subclinical bovine mastitis cases in Baghdad Governorate

Fadwa Abdul Razaq Jameel

APJMBB 33(3): 222-228

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.22

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Mastitis is a serious economic disease that can be caused by fungal pathogens from the genus Candida. This study aimed to isolate and identify Candida parapsillosis from subclinical cases of bovine mastitis and to evaluate its main virulence factors, including proteinase production (using albumin and casein as substrates), haemolytic activity, urease production, and biofilm formation. Between November 2023 and March 2024, fifty milk samples were collected from various locations within Baghdad Governorate and the surrounding suburban farms of Abu-Ghraib. Each milk sample was tested for the California Mastitis Test (CMT). The results showed that 84% of the samples were positive for CMT. Forty-four mycotic isolates were identified, including six isolates of Candida spp. (33.3%), of which two were Candida parapsilosis (4.5%), along with several other fungal species. All of Candida parapsilosis isolates were identified macroscopically by culturing on Sabouraud Dextrose Agar (general medium), CHROM Agar and BIGGY Agar (special, selective and differential media) at 37°C and 25ºC for 3-5 days and microscopically using lactophenol cotton blue stain. This study demonstrated that C. parapsilosis exhibited several virulence factors, including the ability to degrade albumin, hydrolase urea, produce haemolytic activity on Blood Agar and form biofilms, but was unable to degrade casein. The findings indicate that cow milk contains various types of fungi, including molds and yeasts. Importantly, the yeast species identified were not limited to C. albicans but also other Candida species, with a notable prevalence of C. parapasilosis. This species may act as an opportunistic pathogen in bovine mastitis.

Optimization of PCR protocol for SSR marker based genetic diversity assessment of Citrus genus in Nepal

Nabin Narayan Munankarmi, Sujan Chaudhary, Sagar Gautam, Neesha Rana, Tribikram Bhattarai, Sangita Shrestha 

APJMBB 33(3): 229-238

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.23

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The Citrus genus thrives in a wide variety of landscapes in Nepal. Understanding the genetic diversity of these cultivars can be a critical stride towards conservation and sustainable agricultural practices. The present study revolves around the optimization of the PCR protocols for Simple Sequence Repeats (SSR) marker-based genetic diversity assessment with the aim to facilitate the study of understanding the genetics of Citrus species. To achieve the aim, the PCR protocol was checked for the optimum result with the varying concentrations of DNA, MgCl₂, primer, dNTPs, and Taq polymerase and different cycling conditions. The Cetyltrimethyl ammonium bromide (CTAB) extraction method was used to extract DNA from the ground preserved leaves of the selected sample, and the SSR-PCR products were visualized on a 4% agarose at 70 V for one and a half hour. The optimized PCR reaction conditions that gave clear, scorable, and sharp bands were found to be 25 ng DNA, 3.0 mM MgCl₂ , 0.2 mM dNTPs, 0.4 µM primers, and 2 units Taq DNA polymerase, and the best PCR cycling conditions consisted of initial denaturation at 94°C for 5 min followed by 35 cycles of denaturation at 94°C for 30 s, annealing at primer-specific temperature (47-61°C) for 1 minute, elongation at 72°C for 30 s, and final elongation at 72°C for 4 minutes. Finally, Citrus species genotypes were successfully assessed for the genetic diversity using thus optimized SSR-PCR protocol, which will help in finding genetically diverse superior cultivars for off-season production.

The effect of the natural sugar substitute Stevia on biochemical and metabolic markers in rats

Sanjarbek Khabibullaev, Nasrdjan Yuldashev, Nozima Abdullayeva, Zulfiya Ikramova,
Diloza Babakhanova

APJMBB 33(3): 239-244

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.24

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Stevia rebaudiana Bertoni (Stevia) functions as a common natural sugar substitute which has generated interest regarding its metabolic effects on carbohydrates. The research examined stevia's influence on essential biochemical and hormonal indicators through a controlled experimental approach. A total of 25 males white Wistar rats received daily stevia extract doses during a two-month period. The research spanned two months to evaluate glucose metabolism and tolerance through glucose concentration tests as well as glycated hemoglobin and insulin measurements. Blood samples were collected before treatment, on day 30, and on day 60. The glucose tolerance test required glucose administration followed by AUC analysis of glucose concentration throughout the test period. The consumption of stevia resulted in blood glucose elevations reaching 39.5% on day 30 and 66.2% on day 60 while glycated hemoglobin levels increased by 64.7%. The insulin levels increased by 49.3% on day 30 but demonstrated partial stabilization by day 60. The extended consumption of stevia resulted in major increases of liver enzymes (ALT increased by 47.8% at day 30) together with elevated urea and reatinine levels and decreased calcium and potassium concentrations. The results show that long-term stevia consumption results in elevated blood sugar levels and insulin levels and impaired glucose control. The natural sugar substitute reputation of stevia does not protect users from metabolic risks that occur with extended consumption even at recommended dosages.

CRISPR-Cas9 genome editing in microalgae for improved high-value products (HVP) production

Fazleen Haslinda Mohd Hatta, Nurin Nisa’ Ahmad Zamri, Norazlina Ahmad

APJMBB 33(3): 245-261

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.25

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Microalgae are a broad class of photosynthetic, eukaryotic microorganisms that transform carbon dioxide and solar energy into high-value products (HVPs), which have significant commercial value. They are viewed as promising platforms for HVP production. With the global population estimated to reach approximately 9.22 billion by 2075, microalgae are recognised for their resilient and remarkably effective biofactories. However, despite their industrial relevance and environmental advantages over land plants, microalgae-based HVP production requires further optimisation to become commercially viable. Hence, genome editing tools such as clustered regularly interspaced short palindromic repeats/Cas protein 9 (CRISPR-Cas9) are a potential strategy to generate microalgae strains that promote the production of HVP more efficiently to meet industrial demand compared to other genome editing techniques such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). This review paper examines the potential and effectiveness of CRISPR-Cas9 in microalgae for enhancing the production of HVP, particularly PUFA, carotenoids, mycosporine-like amino acids, and vitamins. The literature search used online databases to consider the inclusion and exclusion criteria. In conclusion, due to its effectiveness, CRISPR-Cas9 is recognised as the most widely used genome editing technique for enhancing microalgae HVP production.

Latest advancements in IVF technology in livestock animals

Ala Sai Divya Teja, Raghavendra B N, Sudhakar Kancharla, Prachetha Kolli, Gowtham Mandadapu, Prasana Kumar Rath, Bidyut Prava Mishra, Manoj Kumar Jena

APJMBB 33(3): 262-274

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.26

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Assisted Reproductive Technologies (ARTs), particularly in vitro fertilization (IVF), have revolutionized livestock breeding by enhancing genetic gain, reproductive efficiency, and conservation efforts. This review article includes the recent advancements and applications of IVF across species like cattle, pigs, and horses. It aims to provide a role of complementary techniques like intracytoplasmic sperm injection (ICSI), gamete and zygote intrafallopian transfer (GIFT/ZIFT), cryopreservation, and genomic screening in improving embryo quality and animal reproduction. The integration of IVF with other technologies resulted in various advancements like optimized vitrification protocols, novel sperm capacitation methods and genomic tools to enable precise trait selection for disease resistance, productivity, and adaptability. The combination of IVF with ovum pick-up (OPU) and embryo transfer has enabled high yields in embryos for mass production of livestock animals to tackle the food crisis due to rising human population. Along with these technologies, challenges need to be answered which include polyspermy in porcine IVF, reduced embryo cytotolerance in goats, large offspring syndrome (LOS) linked to serum-based media, and species to species technical barriers. Innovations such as serum-based media, use of embryokines and gene editing tools like CRISPR-Cas9 technology present new opportunities to optimize IVF efficiency and reduce developmental anomalies. Livestock breeding strategies can be integrated with genomics and advanced reproductive technologies to meet global demands for food security, sustainability, and animal welfare. This review article also highlights the transformative impact of IVF in livestock biotechnology and outlines future directions to overcome current challenges and enhance practical application in animal reproductions.

Enhancing snail resistance to pathogens and parasites:
Advances in bioengineering and detection methods

Yevgen Kotukh, Nataliia Lugovska

APJMBB 33(3): 275-281

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.27

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Snails represent valuable sources of nutrition and bioactive compounds for food, feed, and pharmaceutical applications. However, their susceptibility to pathogens and parasites poses significant risks to human and animal health, limiting their commercial potential. This review examines current bioengineering approaches for enhancing snail resistance to infectious agents and evaluates diagnostic methods for pathogen detection. A comprehensive analysis of scientific literature was conducted using systematic review methodology. Key findings indicate that genetic modification techniques, particularly CRISPR/Cas9, TALEN, and zinc finger nucleases, offer promising solutions for developing pathogen-resistant snail populations. Molecular diagnostic methods, including PCR and next-generation sequencing, demonstrate superior sensitivity and accuracy for pathogen detection compared to traditional approaches. The implementation of these biotechnological strategies could significantly improve snail farming safety, reduce reliance on chemical treatments, and enhance the production of high-quality biological materials for various applications including space and military nutrition programs.

Biotechnology and 3D printing in transforming prosthetic limb manufacturing for personalised healthcare

Kai-Xin Chiam, Nancy Choon-Si Ng, Long Chiau Ming, Rebecca Shin-Yee Wong

APJMBB 33(3): 282-288

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.28

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The use of three-dimensional (3D) printing in the manufacturing of prosthetic limbs has become a revolutionary technology that presents new opportunities of customisation, affordability and accessibility. Conventional prosthetic production is usually expensive, labour-intensive, and has a shortage of capacity to meet various anatomical requirements, particularly in low-resource environments. This narrative review examines how biotechnology and additive manufacturing are coming together to develop prosthetic limbs. It combines technological innovations and social and medical consequences, relying on the existing literature and practice. Findings showed that 3D printing allows high levels of customisation by means of digital scanning and modelling, increases affordability by minimising manual labour and material waste, and increases access by means of decentralised, localised production. Such initiatives as the Victoria Hand Project prove its scalability in underserved communities. In addition to the advantages of technology, 3D-printed prostheses increase patient satisfaction, social inclusion, empower users and local healthcare systems. Nevertheless, some obstacles still exist such as material constraints, regulatory uncertainty, ethical issues and standardisation barriers. In conclusion, the 3D printing in prosthetics is a major step towards inclusive and personalised healthcare. In order to achieve its full potential, the future should be aimed at enhancing materials, regulation, and integrating ethical frameworks into the innovation pipeline.

In Silico prediction of multi-epitope vaccine candidates targeting chikungunya virus structural proteins in Malaysian populations

Hanis Nabilah Mohd Nazman, Nurfatin Husna Suhaime, Ewe Seng Ch’ng, Mohammad Syamsul
Reza Harun, Muhammad Amir Yunus, Siew Kit Ng, Siti Aminah Ahmed, Asmida Isa

APJMBB 33(3): 289-300

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.29

Click here to download [PDF] [Supplementary File]

Chikungunya is a re-emerging mosquito-borne disease caused by the chikungunya virus (CHIKV). It typically causes fever, rash, arthralgia, and muscle pain, adversely affecting the quality of life and has socio-economic and health impacts in epidemic areas. Despite decades of efforts, there is no effective treatment or prevention for CHIKV, making it a significant health-related quality of life issue. Hence, this study proposes innovative immunoinformatic and bioinformatics-driven approaches to identify CHIKV antigens as promising targets for multi-epitope vaccine design for the Malaysian population. The analysis revealed a total of 31 epitopes that matched the most common HLA alleles in Malaysian populations. These epitopes met the criteria for multi-epitope vaccine candidates with high antigenicity, non-allergenic, non-toxicity, and sufficient stability characteristics. This study presents promising vaccine candidates with robust design and demonstrated immunogenicity to address chikungunya's global health burden.

Cloning and expression of recombinant outer membrane protein A of Orientia tsutsugamushi using Escherichia coli expression systems

Noor Hanisa Harun, Mohd Yusri Idorus, Jamal Houssaini, Ang-Lim Chua, Sneha Adhavan, Jia Jia Mah, Kelvin Wei Ting Low, Shelby Xuan Lin Lam, Seok Mui Wang

APJMBB 33(3): 301-310

Article DOI: https://doi.org/10.35118/apjmbb.2025.033.3.30

Click here to download [PDF] [Supplementary File]

OmpA of Orientia tsutsugamushi (OtOmpA) plays an important role during multiple stages of scrub typhus pathogenesis. The bacterial expression system which involves cloning, is commonly used for the production of recombinant proteins. The cloning technique requires appropriate optimization and selection of an efficient expression vector. Therefore, this study aims to identify a suitable expression vector for the production of recombinant OtOmpA (rOtOmpA). Three different expression vectors with different properties were selected: pET-14b(+), pET-20b(+) and pET-28a(+). The DNA sequence of OtOmpA was cloned into the vectors and transformed into the TOP10 cloning host, respectively. Positive colonies carrying the recombinant plasmid were identified by using polymerase chain reaction and agarose gel electrophoresis. DNA sequencing was performed to verify the sequence of OtOmpA. Subsequently, each recombinant plasmid was transformed into the BL21(DE3)plySS expression host respectively. The rOtOmpA protein was expressed, extracted and analyzed by SDS-PAGE and Western blot. Analysis with anti-His tag antibodies showed only pET 20b(+) expresses rOtOmpA, which was confirmed by one-dimensional capillary liquid chromatography with tandem mass spectrometry (1D-LCMS/MS). Only pET-20b(+) possessed a pelB leader coding sequence incorporated in the upper region, closer to the ribosome binding site. The presence of the pelB leader may support the production of rOtOmpA, making pET-20b(+) a compatible expression vector. The production of rOtOmpA is crucial for further analysis of its role in the pathogenicity of O. tsutsugamushi.