Volume 34(1); 2026

The CRL4^Cdt2 E3 ubiquitin ligase: A comprehensive review of its substrates, mechanisms, and roles in genomic stability

Afiqah Aimie Murba, Muhammad Aidil Ibrahim, Sarah Shazwani Zakaria, Nurul Hidayah Adenan, Mohamad Ikhwan Jamaludin, Hideo Nishitani, Mohd Shukuri Mohamad Ali, Mu’adz Ahmad Mazian

APJMBB 34(1): 1-22

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.01

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The CRL4^Cdt2 ubiquitin ligase complex plays a pivotal role in maintaining genomic integrity and regulating the cell cycle, specifically during the S phase. By targeting key proteins such as Cdt1, p21, and Set8 for ubiquitination and degradation, CRL4^Cdt2 ensures proper DNA replication and cell cycle progression. Dysregulation of this complex has been implicated in various cancers, including melanoma, breast cancer, gliomas, and ovarian cancer, where elevated levels of Cdc10-dependent transcript 2 (Cdt2) expression is often associated with poor prognosis and increased tumor aggressiveness. CRL4^Cdt2 role extends beyond cell cycle regulation, as it also participates in the DNA damage response by degrading proteins like XPG, which is essential for nucleotide excision repair. Impaired CRL4^Cdt2 function leads to DNA re-replication, genomic instability, and cancer progression. Recent studies have highlighted the therapeutic potential of targeting CRL4^Cdt2 in cancer treatment, with inhibitors like pevonedistat showing promise in preclinical models. However, challenges remain, including the lack of a three-dimensional structure for Cdt2, which limits our understanding of its substrate recognition and degradation mechanisms. This review revisits the role of CRL4^Cdt2 in regulating its cellular substrates, updated substrates targeted by CRL4^Cdt2, explores its pathological consequences in cancer, and discusses potential therapeutic strategies to target this complex, offering new insights into its function and clinical relevance.

Effects of extracellular vesicles of Clostridium butyricum on migration, PANoptosis, and macrophage polarization in lipopolysaccharide-induced RAW264.7 macrophages

Zhang Qingyu, He Kailun, Mohd Shafiq Aazmi, Mohd Fakharul Zaman Raja Yahya

APJMBB 34(1): 23-34

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.02

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Extracellular vesicles (EVs) are nano-sized membrane-bound vesicles released by cells, playing a crucial role in intercellular communication and exchange between cells and their hosts. EVs from probiotics serve as important carriers, enabling probiotics to interact with host cells and exert their beneficial effects. Clostridium butyricum-derived EVs have been shown to exhibit a high yield, contain minimal toxic substances, and possess the ability to modulate cytokine expression in macrophages. This study aimed to investigate the effects of C. butyricum-derived EVs on cytokine expression, migration, PANoptosis, and polarization in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. EVs were isolated from C. butyricum and characterized using transmission electron microscopy and nanoparticle tracking analysis. Quantitative PCR (q-PCR) was employed to assess cytokine expression in RAW264.7 macrophages, while the transwell assay was used to evaluate their migration capacity. Flow cytometry and q-PCR were further utilized to examine the PANoptosis and polarization of the macrophages. The results demonstrated that C. butyricum-derived EVs could promote the expression of proinflammatory cytokines, such as IL-6, IL-12, and TNF-α (p<0.05), while inhibiting anti-inflammatory cytokines like IL-4 and TGF-β (p<0.05). The EVs also restored the migratory ability of RAW264.7 macrophages. Furthermore, C. butyricum-derived EVs exhibited a protective effect on LPS-stimulated RAW264.7 macrophages by reducing PANoptosis through the inhibition of ZBP1 and iNOS gene expression (p<0.05). Additionally, these EVs induced M1 polarization of the macrophages. This study provides novel insights into the immunoregulatory effects of C. butyricum-derived EVs on macrophages, highlighting their potential therapeutic applications.

Unveiling molecular melanogenesis inhibition of lime
(Citrus aurantifolia (Christm.) Swingle) peel extract

Clarissa Nadya Santoso, Brigitta Amanda Maharani, K. Ariex Widyantara, Adam Hermawan,
Rini Dwiastuti, Agustina Setiawati

APJMBB 34(1): 35-45

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.03

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The overproduction of melanin induces multiple types of hyperpigmentation, including post-inflammatory melanoderma, solar lentigo lesions, and melasma. Consequently, the design of melanogenesis inhibitors as skin-enlightening agents through various molecular mechanisms is urgently needed. Citrus aurantifolia (Christm.) Swingle, or lime, has garnered significant interest as a natural product that contains secondary metabolites to inhibit melanogenesis. Nevertheless, the molecular mechanism of its effect on melanogenesis is not yet fully understood. The lime peel was extracted using 70% ethanol, and the major compounds of the extract were detected by thin-layer chromatography (TLC). The gene target and the top ten target proteins associated with molecular pathways were further investigated based on bioinformatics analysis. Additionally, molecular docking studies were carried out on two specific protein targets to assist in further study.  Finally, in vitro study of tyrosinase (TYR) and carbonic anhydrase 2 (CA2) enzyme inhibition was carried out to confirm the extract's anti-melanogenic effect. There are 10 target genes, including CA2, CYP1B1, CA1, CYP19A1, CA9, ABCB1, CA3, ABCG2, TYR, and ADORA1. An in silico study of hesperidin (HD) and tangeretin (TN) to bind with TYR and CA2 found that the binding affinity was binding energy of -10.01 and -6.85 kcal/mol.  The in vitro study demonstrated that the extract inhibited TYR with an IC₅₀ value of 59.71 ppm, which is in vitro more effective in inhibiting it than kojic acid. Based on these findings, Citrus aurantifolia peel is effective in impeding melanogenesis and can be explored as a plant-based cosmetic.

Extract of Allium sativum L. as a sustainable biopesticide candidate: Phytochemical, toxicity, and molecular characterization

Slamet Isworo, Zahra Yulia Putri

APJMBB 34(1): 46-58

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.04

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The excessive use of synthetic pesticides has raised growing concerns about environmental safety, human health risks, and the emergence of pest resistance. In addressing these issues, this study evaluates the biopesticidal potential of Allium sativum L. (garlic) extract through a multidisciplinary approach involving phytochemical screening, toxicity assessment, and molecular identification. Two hypotheses were proposed: (1) that garlic ethanol extract containing alkaloids, saponins, and steroids would exhibit significant bioactivity against Artemia salina with an LC₅₀ below 100 ppm; and (2) that molecular validation using the chloroplast-encoded matK gene would confirm species identity with ≥ 99% sequence similarity to known A. sativum cultivars. Garlic was extracted via maceration using 70 % ethanol, yielding 19.94% crude extract. Phytochemical tests confirmed the presence of alkaloids, saponins, and steroids. Acute toxicity analysis using the Brine Shrimp Lethality Test (BSLT) yielded an LC₅₀ of 93.54 ppm, classifying the extract as moderately toxic by EPA guidelines. Sequencing of matK gene (854 bp) revealed 99.74% similarity to A. sativum cv. Cang ShanPuKe (GenBank: MK335928.1), supported by a 98% bootstrap in phylogenetic analysis. These findings validate both hypotheses and establish garlic extract as a bioactive, molecularly verified candidate for eco-friendly pest control within sustainable agricultural frameworks.

Expression profiling and clinicopathological correlation of miR-100-5p and miR-141-3p in HPV-mediated cervical carcinogenesis

Nur Sabrina Abd Rashid, Ahmad Aizat Abdul Aziz, Sarina Sulong, Mohd Pazudin Ismail,
Nur Asyilla Che Jalil, Daniel Roza Duski, Nazihah Mohd Yunus

APJMBB 34(1): 59-66

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.05

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Cervical cancer is one of the most prevalent malignancies affecting women worldwide, primarily driven by high-risk human papillomavirus (HPV) infection. Recent findings highlight the critical role of non-coding RNAs, particularly microRNAs (miRNAs), in regulating HPV-mediated oncogenic processes. However, the genotype specific miRNA expression patterns in HPV positive cervical cancer remain underexplored. This study aims to profile miRNA dysregulation in HPV-positive cervical cancer and explore their associations with histopathological parameters. miRNA profiling was performed on paired normal adjacent and cancerous cervical tissues using the NanoString nCounter™ Human V3 miRNA Panel to identify differentially expressed miRNAs (DEMs) followed by Taqman RT-qPCR validation in 50 paired cervical cancer. Expression data were normalized using miR-423-3p and miR-25-3p as endogenous controls, and relative quantification was calculated using the 2^{^−ΔΔCT} method. Statistical analyses were employed to determine their association with clinicopathological data. A total of 62 DEMs were identified, comprising 35 upregulated and 27 downregulated miRNAs (FC ≥ 2.0, p < 0.05, FDR < 0.3). RT-qPCR confirmed downregulation of miR-100-5p (p < 0.0001) and upregulation of miR-141-3p (p = 0.0018) in cancerous tissues. miR-100-5p expression were reduced in squamous cell carcinoma, while miR-141-3p was significantly elevated in early-stage cervical cancer. The distinct expression of these miRNAs across tumour subtypes and stages suggests their involvement in early tumorigenic transformation. In conclusion, differential expression of miR-100-5p and miR-141-3p correlates with tumour histopathological subtype, stage, and HPV genotype highlighting their potential as diagnostic and prognostic biomarkers in HPV-associated cervical cancer.

The effect of vinasse in various trophic conditions on the growth rate and metabolite contents of Euglena sp. with CO₂ aeration

Maya Cindiati, Tia Erfianti, Renata Adaranyssa Egistha Putri, Dedy Kurnianto, Eko Agus Suyono

APJMBB 34(1): 67-82

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.06

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Euglena sp. is capable of accumulating valuable metabolites in its biomass, making it useful across various industries. Due to the high cost of production, this research explores the utilization of cost-effective and readily available carbon sources, such as vinasse. This research aimed to determine the effect of vinasse on Euglena sp. cultivation under various trophic conditions with CO₂ aeration on the growth rate and metabolite contents. The study involved measuring cell density daily using spectrophotometry, while biomass and metabolite contents were measured every three days over an 18-day observation period using standard dry-weight and biochemical assay methods. Cell size and morphology were measured at the beginning and end of cultivation. Data were analyzed using one-way ANOVA followed by Duncan’s multiple-range test. The results indicated that the heterotrophic condition yielded the highest growth rate, biomass, carbohydrates, protein, and paramylon contents. Meanwhile, the positive control exhibited the highest lipid and pigment contents. Aspect ratio value showed that the control treatment produced spindle and elongated-shaped cells, while heterotrophic and mixo-heterotrophic conditions resulted in spherical and spindle-shaped cells. Mixotrophic conditions produced a mix of spherical, spindle, and elongated cells. Our findings suggest that vinasse has potential as an alternative medium for cultivating Euglena sp., offering a cost-effective approach for metabolite production.

Association of DPF3 polymorphisms (rs10129954) with male infertility in the Malaysian population - A preliminary study

Wafa Liyana Mohamad Zainol, Ahmad Aizat Abdul Aziz, Nazihah Mohd Yunus, Sarina Sulong,
Adibah Ibrahim, Aziati Azwari Annuar

APJMBB 34(1): 83-88

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.07

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Male infertility is a multifactorial disorder with genetic abnormalities contributing to 15-30% of cases. Numerous genetic polymorphisms have been investigated to understand their role in male infertility, but few studies show significant associations. The Double PHD Fingers 3 (DPF3) gene is essential for chromatin remodelling and histone-to-protamine transition during spermatogenesis. This study aims to investigate the association between DPF3 (rs10129954) polymorphism and male infertility in Malaysians. This case-control study involved 30 infertile and 30 fertile control males. Genomic DNA was extracted, and genotyping was performed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Associations between genotypes and infertility risk were assessed using Fisher’s Exact Test and odds ratios (ORs) with 95% confidence intervals (CI).  The frequencies of CC, CT, and TT genotypes of DPF3 (rs10129954) polymorphism were 3.3%, 36.7%, and 60.0% in infertile males, and 0%, 23.3%, and 76.7% in fertile controls, respectively. The allele frequencies for C and T were 21.7% and 78.3% in infertile males, compared to 11.7% and 88.3% in fertile control males. No significant differences were observed between the frequencies of DPF3 (rs10129954) genotype and allele in infertile and fertile groups (p>0.05). The T allele was associated with a reduced risk of male infertility (OR: 0.478; 95% CI: 0.176-1.297;  p=0.074), differing from studies in Chicago, Japanese and Han Chinese populations, which linked the T allele to an increased risk. Our findings suggest that the T allele of DPF3 (rs10129954) is not significantly associated with male infertility. Larger studies are needed to confirm this observation.

Production of phytase enzyme by Frateuria sp. UMK-PPS3, a phosphate-solubilising bacterium isolated from paddy rhizosphere

Ainihayati Abdul Rahim, Dibasree Hari Krishnan, Nik Fatin Qharanie Nik Mohd Kamaruzaman

APJMBB 34(1): 89-97

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.08

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Frateuria sp. are known for their plant growth-promoting abilities, particularly in phosphate solubilisation and nutrient cycling within the rhizosphere. Phosphate-solubilising bacteria (PSB) can solubilise organic phosphorus compounds into a form available for plant uptake by producing the phytase enzyme, whose activity is influenced by carbon and nitrogen sources. This study was conducted to identify a previously isolated PSB strain from the paddy rhizosphere and to determine the optimal nutrient condition for phytase production. The PSB strain, labelled as UMK-PPS3, was molecularly identified by 16S rRNA gene analysis. Phytase production was assessed by measuring the amount of inorganic phosphate released from sodium phytate using the molybdate-blue method under different nutrient conditions. To determine the optimal enzyme production, the Pikovskaya medium was prepared with varying carbon sources, including glucose, fructose, and sucrose, while maintaining ammonium sulphate as the nitrogen source. Additionally, the production of phytase enzyme under different nitrogen sources, namely ammonium sulphate, potassium nitrate, and sodium nitrate, while maintaining glucose as a carbon source, was also tested. Molecular identification revealed that isolate UMK-PPS3 shared 99% similarity with Frateuria aurantia based on 16S rRNA gene analysis. The results for phytase production showed that the isolate produced the highest phytase activity in the presence of glucose and ammonium sulphate, with 37.620 ± 0.185 U/mL and 33.542 ± 0.731 U/mL, respectively. These findings highlight that determining optimal nutrient conditions can enhance phytase production and provide essential information for designing biofertilizer formulations that sustain the enzyme activity and phosphorus-solubilising efficiency of Frateuria sp. UMK-PPS3 under soil conditions.

Systematic review on Ganoderma lucidum and its nanoparticles in oral cancer: Cytotoxicity, apoptosis and immunomodulation

Goot Heah Khor, Gabriele Ruth Anisah Froemming, Siti Nurhidayah binti Mohd Azhar, Siti Nursyahirah binti Haizan, Shivkanya Fuloria, Neeraj Kumar Fuloria

APJMBB 34(1): 98-113

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.09

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Oral cancer carries high morbidity and mortality, yet although many studies have tested Ganoderma extracts, the evidence remains scattered. This study reviewed six years of literature on Ganoderma lucidum (GL) and its nanoparticles (GLNP) for evaluating the impacts of cytotoxicity, apoptosis and immunomodulation on oral cancer. A total of 67 studies of Science Direct (20), Web of Science (12), Scopus (30) and PubMed (5) were retrieved from 447 articles from the electronic databases.  Among these 33 studies focused on GLNP, demonstrating significant cytotoxic effects against oral cancer cells. Additionally, 60 studies examined GL’s active compounds, particularly triterpenoids and polysaccharides, which exhibited anticancer and immunomodulatory properties. The quality of the included studies was evaluated using a modified risk assessment framework. The results showed that in-vitro studies demonstrated the highest quality, with 66% (40 articles) rated as good, 16% (10 articles) as fair, and 18% (11 articles) as poor. Followed by in-vivo studies with 53% (27 articles) rated as good, 33% (17 articles) as fair, and 14% (7 articles) as poor. In contrast, clinical trials showed the lowest quality, with only 22% (2 articles) rated as good, 33% (3 articles) as fair, and 45% (4 articles) as poor. The combined evidences indicate that GL and GLNP can induce apoptosis, suppress tumour growth, and boost immune responses. Nanoparticle delivery further enhances these effects by improving bioavailability and targeted delivery. However, well-designed studies are still required to confirm their true therapeutic value and clarify their clinical role as adjunct or alternative options for oral cancer.

Metagenomic insights from NGS into water and soil microbiomes surrounding the Temuan Orang Asli communities of Serendah, Malaysia

Aishath Maumoon, Navindra Kumari Palanisamy, Heo Chong Chin, Jamal Houssaini

APJMBB 34(1): 114-126

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.10

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The Orang Asli has a profound knowledge of the forest and its resources which are shaped by the complex microbial communities present in the soil and water sources around their settlements. However, there is a significant gap in the understanding of the environmental microbiome associated with Orang Asli communities. Investigating the diversity and composition of these microbial communities could provide insights into the intricate relationships between the Orang Asli and their surrounding ecosystems. Environmental samples were collected from sites in triplicate across the village. Water quality was assessed using National Water Quality Standard (NWQS). Genomic DNA was extracted from the 3 water and 3 soil samples using a commercial DNA extraction kit and sent for NGS. Findings found that Proteobacteria, Bacteroidota, Firmicutes, and Actinobacteria were the dominant phyla across all water and soil samples specific phyla were significantly more abundant in soil. Soil microbiomes consistently exhibited higher alpha diversity compared to water microbiomes across various metrics. The soil quality across these sites varies considerably but were low overall. All the water samples fell under Class II, indicating "Good" water quality according to the DOE Water Quality Index. In conclusion, key phyla differed significantly between the two environments, likely due to varying conditions. More detailed studies are needed to fully understand how environmental factors shape these microbial communities and their functions in soil and water ecosystems.

Anti-proliferative activity of bioactive sphingolipids from Leishmania donovani against human cervical cancer cells (HeLa) 

Mriganka Mandal, Jishu Mandal, Krishna Das Saha

APJMBB 34(1): 127-134

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.11

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Leishmanial total membrane lipids and Sphingolipid(s) play an immunomodulatory role in the synovial fluid mononuclear cells of patients with rheumatoid arthritis. Additionally, they exhibit an anti-proliferative effect on various malignant cells, such as B16F10, as well as several acute myeloid leukaemia (AML) cells, including those referenced in K562, U937, and HL-60. Based on our previous experiment, we here represent the anti-proliferative nature of Leishmanial Sphingolipid(s) on Human cervical malignant cell HeLa. Sphingolipid fractions were isolated from attenuated Leishmania donovani by the Bligh and Dyer method, then separated into three fractions by Thin layer Chromatographic method (TLC). Three fractions were named LSL–1, LSL–2, and LSL–3. HeLa cells were treated with three fractions separately. Data were collected following the treatment with sphingolipids, applying a dose- and time-dependent approach. Trypan blue exclusion and MTT assay were performed to study the antiproliferative pattern of HeLa cells. It has been observed that LSL–1 demonstrated greater bioactivity and showed potential for anti-proliferation. We have analysed the morphological characteristics and DNA breakdown of cells after treatment with LSL–1 (60 µg/ml) using phase contrast microscopy and fluorescence microscopy. Sphingolipid (LSL–1) treated cells exhibited morphological changes, including cell shrinkage and membrane blebbing due to the externalisation of phosphatidylserine in the outer leaflet of the membrane. DNA fragmentation was verified by colorimetric assay. Analysis of morphological changes using phage contrast and fluorescence microscopy has shown that an apoptotic body-like structure is formed due to cell shrinkage and the externalisation of phosphatidylserine from the cell membrane. Additionally, DNA fragmentation was observed through Acridine orange/Ethidium Bromide (AO/EtBr) staining. The ELISA assay technique demonstrated histones containing fragmented DNA in a dose-dependent manner. Preliminary investigations have found that Leishmanial sphingolipids possess potent anti-proliferative activity against human cervical cancer cells.

Heavy metal cleanup from contaminated soil by phytoremediation: An update

Palivela Pranay Pritham, Tadikonda Sai Siri, Sudhakar Kancharla, Prachetha Kolli, Gowtham Mandadapu, Manoj Kumar Jena

APJMBB 34(1): 135-143

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.12

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Phytoremediation can be defined as a modern method of removing pollutants from water bodies and soil using plants as a means of remediating the environment for a relatively cheaper price while ensuring environmental safety compared to traditional techniques, particularly for soil and water containing heavy metal pollutants. The approach employs various processes such as phytoextraction where pollutants are sequestered in harvestable plant biomass, phytostabilization where movement of pollutants is inhibited, phytovolatilization which refers to the conversion of pollutants to a volatile form, and phytodegradation where plant tissues that contain the pollutants need to be broken down significantly transforming the tissues. Of these methods, phytoextraction has more commercial applicability, especially in regions with diffused pollution. The efficiency of phytoremediation varies according to the bioavailability of heavy metals, soil characteristics, and availability of selective vegetative cover.  Various aspects of phytoremediation of the heavy metals have been discussed in this article with more focus on the methods of phytoremediation. It is essential to select appropriate plants for different methodologies, such as use of high-biomass plants to limit food chain contamination and use of aromatics for non-edible plant by-products. Besides, this review highlights the significance of phytoremediation as a well-suited solution for cleanup of heavy metal contaminated soil and its contribution to a clean and healthier surrounding.

Identification and in-silico analysis of non-synonymous single nucleotide polymorphism (nsSNPs) in the human GH1 gene

Rita Maliza, Alimuddin Tofrizal, Putra Santoso, Bramadi Arya, Diana Fadhilah

APJMBB 34(1): 144-154

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.13

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Non-synonymous SNPs can affect protein structure and function, including GH1, which regulates growth and metabolism. This study aimed to investigate the functional and structural effects of non-synonymous single nucleotide polymorphisms (nsSNPs) in the GH1 gene, focusing on how these variations might impact protein function and stability, and to explore the GH1 gene network using bioinformatics tools. Bioinformatics techniques were employed to analyze specific GH1 gene nsSNPs, namely rs2001345 (T3A/T3P) and rs151263636 (A39V). Various computational tools, including SIFT, PhD-SNP, PROVEAN, PolyPhen-2, and PANTHER, were used to predict the effects of these nsSNPs on protein function, while the I-Mutant 2.0 server assessed the impact on protein stability. GeneMANIA was utilized to explore the GH1 gene network and its connections to other gene categories. The study found that the rs2001345 variations (T3A/T3P) were predicted to be neutral with no significant impact on protein function, whereas the rs151263636 variant (A39V) was predicted to be disease-causing, likely reducing the function of the GH1 protein. Additionally, the I-Mutant 2.0 server predicted a decrease in protein stability for these nsSNPs. GeneMANIA analysis revealed strong links between the GH1 gene and 20 other gene categories. The study indicates that the GH1 gene variation rs151263636 (A39V) may significantly impact protein function and structural stability. Despite its limitations, the study highlights the value of computational analysis in identifying GH1 gene-related disorders and its potential application in drug discovery and personalized medicine development.

Modulation of methylglyoxal-triggered oxidative damage and apoptosis in human osteoblasts by metformin, berberine, and genistein

Alyaa Al-Khateeb, Suhaila Abd. Muid, Gabriele Ruth Anisah Froemming

APJMBB 34(1): 155-165

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.14

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Bone fragility in diabetes is linked to redox imbalance and advanced glycation end products (AGEs). Methylglyoxal (MGO), a reactive carbonyl compound and precursor of AGEs, complicates the pathogenesis of diabetes mellitus and oxidative stress. Metformin lowers the AGEs level. Genistein and berberine decrease MGO levels and inhibit AGE formation. This study investigates the individual and combinatorial effects of metformin, berberine, and genistein on osteoblast-treated MGO in terms of apoptosis and oxidative stress. Human foetal osteoblast cells (hFOB 1.19) were incubated with MGO for 24 hours and treated with metformin, berberine, genistein, and combinations of these compounds. The cells were assessed for apoptosis, AGE levels, oxidative stress markers, and the polymerization status of actin and tubulin fibers. Incubation with metformin, berberine, or genistein alone showed that these compounds significantly reduced apoptotic cells (26.16%, 31.83%, and 41.83%, respectively) compared to MGO alone(p<0.001). Metformin or berberine individual treatment significantly decreased AGEs compared to MGO alone (p< 0.001 for metformin and p< 0.01 for berberine). Individual treatment resulted in lower reactive oxygen species (ROS), with the alliance of genistein and metformin leading to reduced ROS production in comparison to individual treatment. MGO-induced osteoblast cell death was significantly attenuated by metformin, berberine, and genistein. The results of the current study suggest that genistein, berberine, and metformin may serve as potential therapeutic compounds that contribute to the alleviation of bone-related complications resulting from MGO harmfulness.

Educational note on primer design for in-frame cloning

Wei Chong Gan, Yan Kien Lee, Natalie Yong Xin Wong, Hien Fuh Ng, Yun Fong Ngeow

APJMBB 34(1): 166-168

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.15

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Enhancing antimicrobial compound production in Streptomyces sp. VITGV100 using chitosan and its nanoparticles

Madhuri Mukindrao Moon, John Godwin Christopher

APJMBB 34(1): 169-181

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.16

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Streptomyces, a gram-positive bacterium, is renowned for producing a wide array of secondary metabolites with therapeutic potential. This study evaluates the efficacy of chitosan and chitosan nanoparticles (CNPs) as elicitors to enhance antimicrobial compound production by Streptomyces sp. VITGV100 against selected human pathogens, including Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, and Staphylococcus aureus. Chitosan (100 and 200 µg/ml) and CNPs (20, 50, and 100 mg/ml) were added to the growth medium, while a control group received no elicitor. Minimum Inhibitory Concentration (MIC) results from the microdilution method confirmed that both elicitors significantly enhanced antimicrobial compound synthesis. GC-MS analysis was performed on the extracts, and bioactive compounds were validated via molecular docking with target proteins (PDB IDs: 1KZN, 1GSK, 4RLC, and 4URM). Among the tested concentrations, 100 mg/ml CNPs demonstrated the highest activity, showing 70% inhibition of E. coli at 5 µl/ml, while chitosan at 200 µg/ml showed 55% inhibition against S. aureus at 7 µl/ml. GC-MS profiling of CNP-treated extracts revealed 36 unique peaks compared to 16 in the control. The compound Cholest-5-ene-16,22dione,3,26- dihydroxy-, (3β,25R)- exhibited the strongest binding affinity with 4URM in docking studies, indicating potent antimicrobial potential. The results confirm that CNPs are more effective than chitosan in stimulating secondary metabolite production. Overall, Streptomyces sp. VITGV100 is validated as a potent source of bioactive compounds, and nanoparticle elicitation emerges as a promising strategy for enhancing antimicrobial metabolite biosynthesis for potential therapeutic applications.

Evaluation of antioxidant, antibacterial, and antidiabetic activities in vitro of extracts from Abutilon indicum

Van Mai Do, Le Anh Thu Do, Truong Han Le, Hong Phong Ngo, Chi Linh Tran, Van Hung Mai

APJMBB 34(1): 182-192

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.17

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Plant-derived bioactive compounds play a crucial role in developing natural therapeutic agents. However, limited evidence exists regarding the multifunctional properties of Abutilon indicum (A. indicum). This study aimed to evaluate the in vitro antioxidant, antidiabetic, and antibacterial activities of A. indicum extracts to identify their potential health benefits. The study was carried out by extracting leaves and stems of A. indicum, which were then assessed for antioxidant activity using ABTS, DPPH, NO, RP, FRAP, and TAC assays. Antidiabetic potential was evaluated through α-amylase and α-glucosidase inhibitory assays, while antibacterial activity was determined using the agar well diffusion method against six pathogenic bacteria. The results showed that the leaf extracts of A. indicum exhibited antioxidant activity with IC₅₀ values and in vitro antidiabetic activity with IC₅₀ values, outperforming extracts from roots and stems. For antibacterial activity, leaf extracts strongly inhibited Gram-positive bacteria with MIC values from 250 to 500 µg/mL, while MIC values for Gram-negative bacteria ranged from 1000 to 2000 µg/mL. A. indicum is a promising natural resource for pharmaceutical and health-protective products with antioxidant, antibacterial, and antidiabetic properties. These findings support further exploration for therapeutic applications, although additional in vivo and mechanistic studies are required to validate and extend these results.

Trends in genetics and genomics publications in Malaysia:
A Web of Science bibliometrics review

Amin Abdurrahman Abdul Rashid, Wan Amirul Syazwan Wan Ahmad Munawar, Marjanu Hikmah Elias, Sarina Sulong

APJMBB 34(1): 193-206

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.18

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Genetic research, originating from foundational discoveries in DNA structure and function, has evolved into a pivotal scientific discipline. As sequencing technology becomes increasingly accessible, the field is rapidly entering the 'genomic age,' with Malaysia making significant and distinct contributions through advancements in tropical biodiversity and molecular studies. The rapid accumulation of high-throughput genomic and genetic data highlights the critical need for systematic quantitative analyses to chart scientific output and policy relevance within key geographical regions. In this study, we aimed to analyse the trends in genomic and genetic research related to Malaysia. Records were retrieved from the Web of Science Core Collection using a strategic search approach with terms such as “genetic” and “Malaysia”. The R package, Bibliometrix, and VOSviewer were used to analyse the articles' metadata, including publication trends, contributing institutions, journal sources, authors, and keywords. Thematic analysis was conducted to identify research themes from common words in titles, abstracts, and keywords. A total of 4630 articles published between 1976 and 28 April 2025 were retrieved. The earliest publication was in 1976, and the highest number of publications was recorded in 2022. The thematic analysis revealed that the articles could be categorised into four main research themes: ‘tropical medicine and infectious diseases’, ‘medical genetics and heredity’, ‘biotechnology and agriculture’, and ‘ecology, environment, and preservation’. These findings highlight Malaysia's remarkable advancements in genetic and genomic research, demonstrating the country’s growing expertise in biotechnology and molecular biology. As Malaysia continues to address emerging scientific challenges, it is establishing itself as a key player in applying molecular biology technologies.

Interleukin – 4 (IL-4) gene polymorphism (C-589T) in pregnant women with preeclampsia

Awatif Zabel Bdiwi, Sally Salih Jumaa, Azhar Hamid Rasool, Hayder Hussein Jalood

APJMBB 34(1): 207-213

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.19

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Preeclampsia is a common complication of pregnancy, contributing significantly to maternal mortality worldwide. The C-589T polymorphism in the IL-4 gene is believed to play a role in the impaired immune response associated with gestational hypertension, but this relationship has not been adequately investigated in pregnant women. This study aims to reveal the relationship between IL-4 gene polymorphism (C-589T) and pregnant women with hypertension. The polymorphism (C-589T) in the IL-4 gene is associated with lower blood IL-4 levels in pregnant women with hypertension, which may contribute to an immune dysregulation that increases the risk of preeclampsia. Sequence analysis is used to determine the genotypes and allele frequencies of the IL-4 gene polymorphism. ELISA is used to estimate interleukin-4 in blood serum. The CT genotype is more common in pregnant women with hypertension (30%) compared to the control group (20%). The higher frequency of the TT genotype in pregnant women with hypertension compared to healthy controls may indicate a protective role for the T allele. The present study indicates that the serum IL-4 concentration in pregnant women with hypertension (21.82 ± 3.79 rg/ml) is significantly lower than in the control group (24.03 ± 3.03). The protective trend observed for the T allele and TT genotype warrants further investigation in other studies. Although no strong genetic association was detected, the findings support the hypothesis that immune dysregulation, including lower IL-4 activity, contributes to the development of hypertensive disorders in pregnancy.

Hypoxia-conditioned MSC secretome attenuates CD38 expression and cellular senescence in a diabetic rat brain model

Bhirau Wilaksono, Agung Putra, Chodidjah, Mas Rizky A.A. Syamsunarno, Prasetyowati Subchan, Titiek Sumarawati, Nurul Hidayah, Mohammad Ariq Nazar

APJMBB 34(1): 214-223

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.20

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Diabetes accelerates neurodegeneration, partly through CD38-mediated NAD+depletion and cellular senescence. Secretome hypoxia mesenchymal stem cells (S-HMSCs) secrete anti-inflammatory factors such as IL-10, which may counteract neurodegenerative damage. This study evaluated the effects of S-HMSC therapy on CD38, p16, p21, and SIRT1 expression in the brains of STZ-induced diabetic rats. Male Wistar rats were divided into four groups: SHAM (healthy control), C (diabetic control), P1 (500 µL S-HMSC), and P2 (1000 µL S-HMSC). Diabetes was induced using streptozotocin (65 mg/kg, IP), confirmed by blood glucose >200 mg/dL. S-HMSCs were administered intraperitoneally on days 0, 7, 14, and 21. CD38 expression was assessed using immunohistochemistry (IHC); p16, p21, and SIRT1 were analyzed by qRT-PCR. CD38 expression significantly decreased in P1 (12.72 ± 2.65) and P2 (8.43 ± 1.01) compared to C (19.07 ± 2.70). p16 and p21 expression levels were significantly reduced in P1 and P2 compared to C (P < 0.05). SIRT1 was significantly upregulated in P1 (2.48) but downregulated in P2 (0.67), suggesting a nonlinear dose response. S-HMSC therapy reduces CD38 expression and senescence markers (p16, p21) in the diabetic brain. However, SIRT1 expression exhibits a biphasic dose response. These findings support the potential of S-HMSC therapy for modulating neurodegeneration in diabetes.

Isolation and characterization of non-symbiotic N-fixing bacteria from rhizosphere of Calopogonium mucunoides Desv.

Sri Anggreni Lindawati, Ni Nyoman Sista Jayasanti, I Wayan Suarna, Ni Nyoman Suryani

APJMBB 34(1): 224-233

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.1.21

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Calopogonium mucunoides is a leguminous plant that thrives in low-fertility soils and possesses the ability to fix atmospheric nitrogen. Furthermore, the growth of C. mucunoides is supported by the interactions of rhizospheric bacteria and plant roots. The rhizosphere of C. mucunoides consists of diverse bacteria that are able to provide nutrients like nitrogen and produce plant growth hormones. The nitrogen fixation process in the rhizosphere of C. mucunoides can be carried out by non-symbiotic nitrogen fixing bacteria. This study aimed to isolate non-symbiotic N-fixing bacteria from the rhizosphere of C. mucunoides and evaluate their ability to produce Indole-3-Acetic Acid (IAA) as one of plant growth-promoting bacteria (PGPB) traits. Colony and cell morphology, phenotypic characterisation, IAA assay, 16S rRNA sequencing and phylogenetic analyses were conducted. Nine isolates capable of growth on Ashby’s Mannitol Agar were obtained. All isolates were catalase positive, and had optimal pH and temperature at 7 and 37 ⁰C. Seven of the nine isolates were subjected to 16S rRNA gene analysis and identified as Arthrobacter sp. (isolates K2.1 and K2.2), Ensifer adhaerens (isolate K4), Arthrobacter enclensis (isolate K5), Bacillus sp. (isolate K6), Gordonia namibiensis (isolate K8), and Kocuria sp. (isolate K9). IAA results showed that two isolates, namely K4 and K6 produced significant pink coloration and had similarity with Ensifer adhaerens PZS_S05 (99.78% identity) and Bacillus sp. (100% identity). These results indicated that the isolates from this study possess the potential to act as PGPB.

Volume 34(2); 2026

Antibacterial activity of ethyl acetate extract from Lasiodiplodia pseudotheobromae IBRL OS-64 against Vibrio species in aquaculture, with emphasis on V. parahaemolyticus

Mohd Taufiq Mat Jalil, Mohd Fakharul Zaman Raja Yahya, Nurul Aili Zakaria, Sharifah Aminah Syed Mohamad, Kharul Azmi Mu’azzam Abdul Rahman, Darah Ibrahim

APJMBB 34(2): 1-11

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.01

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Endophytic fungi are known to produce bioactive compounds similar to those of their host plants, making them promising sources of novel antimicrobial agents. The use of endophytic fungi offers several advantages, including reduced dependence on rare plants, sustainable production, lower environmental impact, and rapid metabolite yield. This study aimed to evaluate the antibacterial activity of the endophytic fungus Lasiodiplodia pseudotheobromae IBRL OS-64 against selected aquaculture-related pathogenic bacteria. The antibacterial activity of the ethyl acetate extract of L. pseudotheobromae IBRL OS-64 was assessed against four Vibrio species, namely Vibrio parahaemolyticus ATCC 17802, V. alginolyticus CCB-PB317, V. owensii CCB-PG2, and V. azureus CCB-ST2H16, using disc diffusion and broth microdilution assays. The extract exhibited notable antibacterial activity, with inhibition zone diameters ranging from 9.2 ± 0.4 to 12.4 ± 0.4 mm. Minimum inhibitory concentration (MIC) values ranged from 500 to 1000 µg/mL, while minimum bactericidal concentration (MBC) values ranged from 500 to 4000 µg/mL. Time-kill analysis demonstrated that the extract exerted bactericidal effects at higher concentrations, with bacterial reduction influenced by both extract concentration and exposure duration. Scanning electron microscopy revealed severe structural damage to treated bacterial cells, including membrane disruption, cavity formation, cell crumpling, and lysis. These findings suggest that the ethyl acetate extract of L. pseudotheobromae IBRL OS-64 has potential as an alternative antibacterial agent for controlling bacterial infections in aquaculture products.

Navigating key mutations in RBD and SARS-CoV-2 epitope classification: Enhancing antibody therapeutic development amidst evolving challenges

Kelven Alexsius, Indah Mohd Amin, Norfatimah Mohamed Yunus, Rabiatul Basria S.M.N. Mydin, Farizan Aris, Nurul Aili Zakaria

APJMBB 34(2): 12-35

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.02

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Mutations within the receptor‑binding domain (RBD) of SARS‑CoV‑2 present significant challenges to the development of vaccines and antibody‑based therapies. These alterations enable the virus to evade neutralizing antibodies produced by vaccination and also therapeutic antibodies currently in use. Research has further demonstrated that the RBD plays a critical role in mediating interactions with host cells and can alternate between two conformational states, referred to as “up” and “down.” In light of these challenges and the urgent need for effective countermeasures against emerging variants, the characterization and design of novel antibody formats are essential. This study aims to examine how RBD mutations and epitope classification can inform the advancement of antibody therapeutics, particularly in the context of ongoing viral evolution. Based on the data analysis, key mutations such as E484 and N501, together with the conserved epitope sequence ⁴⁶⁹STEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYR⁵⁰⁹, are proposed as promising targets for the development of antibody‑based therapies and vaccine strategies.

Theophylline-enhanced sperm preparation for assisted reproductive technology

Ali Abbas-Kadhim, Zaid Osama-Ibraheem, Hayder A. L. Mossa

APJMBB 34(2): 36-44

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.03

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Asthenozoospermia is a form of male infertility characterised by a marked reduction in sperm motility. This condition necessitates the use of Assisted Reproductive Technology techniques to achieve successful conception. These techniques require the preparation of semen samples enriched with active and motile sperm. Various preparation methods have been proposed, including density gradient centrifugation (DGC) and swim-up techniques. DGC is more preferred for poor-quality samples. This study aimed to improve the efficacy of the DGC technique using theophylline as an enhancement factor to increase the yield of active motile sperm. Briefly, semen samples from normozoospermic individuals (n=60) and asthenozoospermic individuals (n=80) were collected and analysed using the conventional sperm analysis. Each sample was divided into three equal aliquots, which were exposed to the conventional density gradient centrifugation, incubation with theophylline and to a combination of both treatments. The results showed that theophylline addition improved the yield of active motile sperm, particularly in samples from asthenozoospermic individuals. Overall, the study suggests that theophylline may serve as an effective enhancement factor for isolating and concentrating active motile sperms in semen samples obtained asthenozoospermic males.

Computational insights into the immunomodulatory potential of alizarin: A molecular docking and dynamics study

Sukma Sahadewa, Fara Disa Durry, Olivia Herliani, Mochammad Aqilah Herdiansyah

APJMBB 34(2): 45-55

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.04

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Alizarin (1,2-dihydroxy-9,10-anthraquinone) is a naturally occurring anthraquinone derivative characterized by three conjugated aromatic rings and two central ketone groups. Many studies have also reported the potential of alizarin as an antitumor, anticancer, anti-inflammatory, and antibacterial. This study employs molecular docking simulations to evaluate the potential of alizarin as a natural immunomodulatory agent by assessing its interactions with key immune-related receptor proteins. The receptor proteins used are SYK (PDB ID: 6V0V), SRC (PDB ID: 2BDF), PIK3CG (PDB ID: 7JWE), and ARG1 (PDB ID: 3E6K). Molecular docking was performed using PyRx v.0.8 software, while molecular dynamics validation was conducted using CABS-flex. The results showed that alizarin compounds have drug-like properties with a bioavailability score of 0.55 and comply with the Lipinski Rule of Five. The results of the docking experiments demonstrated that each receptor has the capacity to bind with the compound, as indicated by the binding affinity values found in each complex. Of the four receptor-ligand complexes, alizarin-PIK3CG had the highest binding affinity (1.49 × 10^-7 M), followed by alizarin-SRC (2.49 × 10^-7 M), alizarin-ARG1 (2.21 × 10^-6 M), and alizarin-SYK (1.02 × 10^-5 M). Molecular dynamics validation using CABS-flex showed that all receptors had an RMSF value <3 Å, indicating structural stability and low solubility in the body. ADMET data confirmed that alizarin compounds are non-toxic and water-soluble. These findings suggest that alizarin exhibits promising immunomodulatory potential, particularly through its interaction with PIK3CG. Further in vitro and in vivo studies are warranted to validate its efficacy and therapeutic applicability. 

Genetic characterization of Plectranthus amboinicus (Lour.) using morphology and ITS sequences across eight Vietnamese provinces

Van Duong Thieu, Nam Long Thieu, Van The Tran, Van Hung Mai

APJMBB 34(2): 56-61

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.05

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Plectranthus amboinicus (Lour.) is a spice and medicinal vegetable with tenacious vitality, easy to grow, and able to thrive even in the shade, making it widely cultivated in Vietnam. This study aimed to evaluate the agronomic characteristics of 80 samples of Plectranthus amboinicus (Lour.) from 8 provinces, based on a morphological assessment of stem height, root length, and leaves, combined with molecular biology methods to determine genetic relationships using the Internal Transcribed Spacer (ITS) gene region sequencing. The results of the morphological characteristics of the plant samples showed variations in stem height, leaf length, leaf width, and root length; however, flowers and fruits showed little variation among samples. Samples collected from An Giang and Hau Giang provinces were relatively larger than those from other provinces. Phylogenetic analysis demonstrated a close genetic relatedness among Plectranthus amboinicus (Lour.) genotypes from the eight provinces, while exhibiting a genetic distance of 0.2970 from the control species, Mentha arvensis, an exotic species. Therefore, there were differences in the morphological characteristics of Plectranthus amboinicus (Lour.) among the samples. The amplified sequence of the ITS gene region of the experimental samples was approximately 600 bp in size, and sequencing results showed high similarity to to published Plectranthus amboinicus (Lour.) reference sequences.

Role of A1 and A2 bovine milk in human health and diseases:
An in-depth review

Shivam K. Dubey, Abhishek Thakur, Manoj Kumar Jena, Sudhakar Kancharla, Prachetha Kolli, Gowtham Mandadapu, Sudarshan Kumar, Ashok Kumar Mohanty

APJMBB 34(2): 62-76

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.06

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Milk protein composition has significant effect on the milk quality and nutrient value. Various cattle breeds show distinct milk protein polymorphisms as a result of single nucleotide polymorphism, insertion and deletion of nucleotides, and post-translational modification. The milk casein proteins comprise of one third of the total milk protein content and various variants predominate in different cattle populations. The A1 and A2 alleles of β- casein variants have shown some difference in their effect on human health. A single amino acid difference at 67th position (proline in A2 and histidine in A1) has shown profound effects on their physicochemical and functional properties as well as their effects on human health. The 7-amino acid peptide BCM-7 is produced upon enzymatic cleavage of A1 variant and it is a µ-opioid receptor agonist. The BCM-7 is found to have adverse health effects on the gastrointestinal, cardiovascular, endocrine, and nervous systems. The present review discusses in detail about the A1 and A2 bovine milk and their effects on human health, and how BCM-7 plays role in causing diseases of various organ systems. Future studies should be designed to provide clear guidance on the preferential use of A2 milk and to evaluate any potential consequences of gradually eliminating the A1 β- casein variant from cattle populations through selective breeding. This review presents a comprehensive mechanistic overview of β-casomorphin-7 (BCM-7), with particular emphasis on its interaction with μ-opioid receptors and associated systemic effects. It also discusses methods for the identification of A1 and A2 milk, compares their impacts on human health, and examines the biological effects of BCM-7 across multiple organ systems, including the gastrointestinal, cardiovascular, neurological, and endocrine systems. Furthermore, the review integrates evidence from animal, in vitro, and human studies, alongside considerations of casein genetic variants and dairy breeding strategies. The present review article will guide the researchers for a better research strategy to unravel the details of A1 vs. A2 milk consumption in a long run.

Immunotherapy strategy for cervical cancer targeting HPV E6/E7: Hopes and challenges

Henny Saraswati, Riza Arief Putranto, Mira Asmirajanti

APJMBB 34(2): 77-90

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.07

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Cervical cancer is one of the leading causes of cancer-related mortality in women worldwide. Persistent infection with Human Papillomavirus (HPPV) is recognized as the predominant etiological factor, primarily through the expression of the viral oncoproteins E6 and E7. These proteins disrupt key cell cycle regulatory pathways, promote uncontrolled cellular proliferation, and drive the malignant transformation. Although various therapeutic approaches, including surgery, chemotherapy, and radiotherapy, have been developed to reduce cervical cancer mortality, the limitations of conventional treatments, including toxicity, recurrence, and reduced efficacy in advanced-stage disease, underscore the urgent need for more effective and targeted strategies. In this context, the E6 and E7 oncoproteins have emerged as highly promising therapeutic targets, not only because of their indispensable roles in HPV-mediated oncogenesis but also due to their strong immunogenic properties. Consequently, substantial efforts have been directed toward developing E6/E7-based therapeutic vaccines. Multiple vaccine platforms, including DNA and protein-based vaccines, have demonstrated favorable safety profiles and the capacity to induce robust immune responses. Furthermore, preclinical and clinical studies suggest their potential to generate antitumor activity while offering prophylactic benefits in certain settings. Despite these advances, several challenges continue to limit the clinical translation and therapeutic efficacy of E6/E7-targeted vaccines. One major barrier is the tumor microenvironment (TME), which suppresses host antitumor immunity and attenuates vaccine-induced immune responses. In addition, intratumoral heterogeneity may facilitate immune escape mechanisms, allowing malignant cells to evade immune surveillance and persist, despite treatment. Therefore, future directions for E6/E7-based vaccine development should focus on overcoming these barriers through combination strategies with other immunotherapeutic modalities, particularly immune checkpoint inhibitors (ICIs), as well as the incorporation of more potent adjuvants, multi-epitope vaccine design, and precision medicine approaches. Collectively, these advances may enhance therapeutic efficacy and pave the way for more personalized and durable treatment options for patients with cervical cancer.

Antioxidant and probiotic potential of Lacticaseibacillus paracasei isolates from Malaysian water kefir grains

Muhammad Asyraaf Haja Maideen, Nurulain Syahirah Razali, Nor Farahin Azizi, Noorshafadzilah Talib,
Nik Mohd. Afizan Nik Abd. Rahman, Mohd. Azuraidi Osman, Suraini Abd-Aziz, Noorjahan Banu
Alitheen

APJMBB 34(2): 91-102

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.08

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Water kefir is a traditional fermented beverage that comprises a microbially diverse community enriched in lactic acid bacteria (LAB). Previous studies have examined the predominant species recovered from each sample. However, functional strain-level variation among multiple LAB isolates within a single kefir grain source remains insufficiently explored. This study aimed to isolate and characterise Lactobacillus species from Malaysian water kefir grains and evaluate their antioxidant activity and probiotic potential. Bacterial isolates were initially characterized through colony morphology, Gram staining, and catalase testing. Antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. Isolates demonstrating high levels of antioxidant activity were evaluated for probiotic characteristics, including acid and bile salt tolerance, adhesion to intestinal epithelial cells, and antibiotic susceptibility. Molecular identification was performed through 16S rRNA gene sequencing. Among fifteen isolates, isolates 2 and 5 exhibited significantly higher antioxidant activities (p < 0.05) with ABTS radical scavenging activities of 72.70 ± 3.38% and 77.74 ± 3.27%, and DPPH radical scavenging activities of 41.62 ± 1.18% and 48.07 ± 5.18%, respectively. Probiotic characterization demonstrated that isolate 2 exhibited higher bile tolerance and adhesion capacity with a 93.57 ± 1.31% survival rate under bile salt stress and 88.8 ± 6.3% adhesion rate to HT-29 cells. Molecular identification identified both isolates as Lacticaseibacillus paracasei, a species widely associated with probiotic functionality. Taken together, these findings highlight isolate 2 as a promising probiotic candidate with antioxidant potential, warranting further investigation to validate its functional efficacy and safety.

Computational insights into andrographolide and its possible role in prostate cancer

Umi Baroroh, Azwar Faturochman, Melvia Sundalian, Dewi Astriany, Komar Ruslan

APJMBB 34(2): 103-114

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.09

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Prostate cancer remains a major global health concern, highlighting the need for continuous investigation of bioactive compounds with potential relevance to its management. This study presents a computational assessment of andrographolide through an integrated approach combining Absorption, Distribution, Metabolism, and Excretion (ADME) prediction, network analysis, molecular docking, and molecular dynamics (MD) simulation. ADME profiling indicated that andrographolide meets drug-likeness parameters, including Lipinski’s Rule of Five. Network analysis of prostate cancer-associated genes using the STRING database revealed a cluster of six nodes, from which three key targets, androgen receptor (AR), CYP17A1, and SRD5A2, were selected for docking analysis. Docking results suggested favorable interactions between andrographolide and these targets. Furthermore, MD simulations of the androgen receptor complex, with metribolone as a reference ligand, demonstrated stable binding patterns and comparable binding energies (-49.95 and -50.94 kcal/mol, respectively). While further experimental validation is required, these computational findings provide preliminary insights into the possible role of andrographolide in prostate cancer research.

Antibiotic resistance patterns and vanA gene detection in Staphylococcus aureus clinical isolates from Gunung Jati Hospital, Cirebon, Indonesia

Maikel Ntolidi, Stalis Norma Ethica, Maya Dian Rakhmawatie

APJMBB 34(2): 115-123

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.10

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The emergence of methicillin-resistant Staphylococcus aureus (MRSA) contributes to increased morbidity and mortality. Vancomycin is a last-resort antibiotic for the treatment of MRSA infections and its extensive use may lead to an increased occurrence of vancomycin-resistant S. aureus (VRSA). Detection of the vanA gene can be used to understand the mechanism of vancomycin resistance in S. aureus and to provide epidemiological data for preventing the spread of resistant strains. This study aimed to determine antibiotic resistance patterns and detect the presence of the vanA gene in S. aureus clinical isolates obtained from patients at Gunung Jati Hospital, Cirebon, Indonesia. A total of twenty S. aureus isolates were identified, and antibiotic susceptibility testing was performed using the Vitek‑2 Compact system. Detection of the vanA gene was carried out by polymerase chain reaction analysis. The Vitek-2 Compact analysis showed that 9 isolates (45%) were identified as MRSA, 2 isolates (10%) as VRSA, 1 isolate (5%) showed characteristics of both MRSA and VRSA, and 8 isolates (40%) were methicillin-sensitive S. aureus (MSSA). Differences in electrophoresis banding patterns were observed among VRSA, MRSA, and MSSA isolates. However, the vanA gene was not detected in any of the S. aureus clinical isolates from Gunung Jati Hospital. Although the vanA gene responsible for vancomycin resistance was not detected, the clinical isolates demonstrated resistance to multiple antibiotics. Therefore, increased vigilance in antibiotic use is necessary. Implementation of antibiotic stewardship strategies, including the use of sensitive antibiotics, may help reduce the risk of disease severity due to infection.

Effect of explant types and medium treatment on Clinacanthus
nutans callus induction with secondary metabolite analysis

Saiyidah Nafisah Hashim, Siti Mazleena Mohamed, Tay Chia Chay, Pathimah Abdol Latif, Putri Shareen
Rosman, Mohammad Feizal Daud, Norrizah Jaafar Sidik

APJMBB 34(2): 124-134

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.11

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The medicinal properties of Clinacanthus nutans are mostly ascribed to its abundance of bioactive compounds. Therefore, the establishment of C. nutans callus culture is anticipated to play an integral part in the sustainable mass production of these beneficial compounds. The study used Murashige and Skoog (MS) medium added with the hormones of Naphthalene Acetic Acid (NAA), 6-Benzylaminopurine (BAP), or wood vinegar (WV) in single and combination methods. The types of explants applied for this callus induction were nodes and leaf explants. The resulting callus was then further multiplied on MS medium with 0.5 mg/L NAA. Within 7 weeks, measurements on the total phenolic content (TPC), total flavonoid content (TFC), and antioxidant capacity were conducted, and the results were subjected to correlation analysis. The MS + 0.5 mg/L NAA (93.11%) was the most effective medium for inducing callus in C. nutans from nodal explants. The callus extract TPC, TFC, DPPH, and FRAP analyses values were determined at the highest in week 7, given 185.98 mg GAE/g extract, 9.75 mg QE/g extract, 55.72 %, and 32.20 g FeSO₄/g DW in C. nutans callus, respectively. The positive correlation between TPC and TFC with the antioxidant activities was documented.

Peperomia pellucida mitigates glomerulotubular and interstitial damage following cigarette smoke exposure in Wistar rats

Yudha Dwi Suryanto, Johanes Aprilius Falerio Kristijanto, Nur Khamidah, Budiono Raharjo, Aryagani Manggala Athaseno, Farida Anggraini Soetedjo

APJMBB 34(2): 135-142

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.12

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Secondhand smoke (SHS) exposure injures renal tissue and may accelerate chronic kidney disease, highlighting the need for accessible nephroprotective strategies such as Peperomia pellucida (PP). We conducted a true experimental, post-test–only study in male Wistar rats (n = 21; 8–12 weeks; 180–220 g) randomized to three groups (n = 7 each): normal control, SHS, and SHS plus PP. PP extract (400 mg/kg/day, oral gavage) was administered for five weeks; SHS exposure (one cigarette/rat/day, whole-body chamber) was delivered for four weeks. At endpoint, kidneys were processed for hematoxylin–eosin staining, and histopathology was scored semiquantitatively across endothelial, glomerular, tubular, and interstitial compartments. Two-way ANOVA with Tukey’s post hoc test (α = 0.05) evaluated group differences. Overall effects were significant for all compartments (p < 0.0001). SHS produced the highest injury burden, indicating diffuse renal damage with a tubulointerstitial predominance. Co-treatment with PP significantly reduced lesion scores versus SHS alone across all compartments (p < 0.05), although values did not fully normalize to the control group, indicating partial structural preservation rather than complete rescue. These results demonstrate that PP confers measurable nephroprotection against SHS-induced renal injury in vivo, most notably attenuating tubulointerstitial lesions. Future work incorporating mechanistic biomarkers, dose–response optimization, and pharmacokinetic profiling is warranted to define the mode of action and enhance translational potential.

Glucose and insulin modulate inflammation and glycolipid metabolism genes in goose muscle via IGFBP2 during fatty liver development

Fangbo Li, Muhammad Syahmi Hishamuddin, Shiou Yih Lee, Bashir Jarrar, Chun Hung Chang, Tuoyu Geng

APJMBB 34(2): 143-154

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.13

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Muscle, the largest tissue in geese, is crucial for developing goose fatty liver. Investigating the influence of Insulin-like Growth Factor Binding Protein 2 (IGFBP2) on muscle during fatty liver development is crucial for understanding its regulatory mechanisms. In this study, twenty-four 70-day-old male Landes geese were divided into two groups: an overfeeding group and a control group (n = 12 each). Pectoral muscle samples were collected at 82 and 94 days of age. Additionally, twenty-four 10-day-old geese were divided into a glucose injection group and a saline control group (n = 12 each), with pectoral muscle samples collected 2 hours post-injection at 17 days of age. Goose primary myocytes were treated with glucose (0, 25, 50, and 100 mmol/L) or insulin (0, 5, 10, and 20 nmol/L) and transfected with an IGFBP2 overexpression vector or an empty vector. Results demonstrated that overfeeding, glucose injection, 50 and 200 mmol/L glucose treatment, and 10 and 20 nmol/L insulin treatment significantly inhibited the expression of IGFBP2 in both muscle and cell samples (P < 0.05). Overexpression of IGFBP2 increased the expression of LOC106030908, FGB, and LOC106040417, while inhibiting PLPP4, PTGS2, IL6, and LOC106041919 (P < 0.05). Notably, gene expression showed an inverse pattern in the muscles of overfed geese. Additionally, glucose and insulin upregulated LOC106041919 and LOC106040417 while downregulating LOC106030908 in goose myocytes (P < 0.05). These results suggest that glucose and insulin modulate inflammation and glycolipid metabolism genes in goose muscle via IGFBP2 during fatty liver development, providing new molecular insights.

Population-specific pathogenic variants in blood disorder genes among Orang Asli and Malay populations: Whole-genome insights with 3D modelling, functional prediction, and interaction mapping

Aishah Farliani Shirat, Nurul Azmir Amir Hashim, Mohd Nur Fakhruzzaman Noorizhab, Eng Keng Seow, Mohammad Masrin Md Zahrin, Thane Moze Darumalinggam, Teh Lay Kek, Mohd Zaki Salleh

APJMBB 34(2): 155-167

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.14

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This study aimed to identify genetic variants associated with blood disorders in the Orang Asli and Malay populations by analyzing previously sequenced whole genomes thereby shedding light on the genetic burden within these groups. We focused on 14 key genes: BMP2, CD164, CYBRD1, EPAS1, EPO, HAMP, HBB, HFE, MTHFR, SH2B3, SLC40A1, TF, TMPRSS6, and VHL, chosen for their roles in hematopoiesis, iron metabolism, erythropoiesis, and oxygen homeostasis which are essential factors in blood disorders. We developed a bioinformatics pipeline to mine whole-genome sequences and map variants to public databases, identifying pathogenic SNPs linked to blood disorder risks. We predicted the functional impact of these SNPs using SIFT and PolyPhen-2. Of the 4,535-blood disorder-related nsSNPs identified, 45 were found in the Orang Asli and Malay populations. We further analyzed these variants for functional impact, conservation, and stability using HOPE, MutPred2, and I-Mutant, with Jalview for stability evaluation. Among the identified variants, rs235768 (BMP2), rs1799945 (HFE), rs1801133 (MTHFR), rs41298977 (TF), and rs190329416 (TMPRSS6), were classified as pathogenic. Compared with global population data, we observed significantly higher allele frequencies of rs1799945 and rs1801133 in both populations, indicating a population-specific risk for iron overload and folate metabolism disorders. These mutations impact protein interfaces and allosteric sites, influencing cell proliferation, iron transport, and homeostasis. In conclusion, the five identified variants are significant pathogenic factors that increase the risk of blood disorders in the Orang Asli and Malay populations. Further research is necessary to clarify their roles and implications.

Turbinado sugar in perspective: An evidence-based review on myths, facts, and nutritional significance

Mary Nancy Flora Rayappan, Nancy William, Kuppuswami Govindan, Johnson Irudayam, Ramya Ravi, Ramalatha Mariyappan

APJMBB 34(2): 168-181

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.15

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Turbinado sugar is often placed as healthier, natural white sugar alternative, based on the minimal processing and maintenance of trace molasses content. The scientific rationale behind such assertions is critically evaluated in this review, by taking into account the nutritional, metabolic, environmental, and ethical aspects of turbinado sugar production and utilization. Available evidence suggests that turbinado sugar and white sugar are very comparable in caloric content and glycemic load, and there are no notable health benefits. Further, most purported health benefits of turbinado sugar are marketing and publicly led misconceptions and not scientifically backed. The environmental issues of water usage, soil loss, and carbon emissions are also relevant to turbinado sugar, and sustainability is place-specific farming and processing procedures. Ethical considerations of exploitation of labor and greenwashing in labeling further muddy the waters. The review is urging enhanced nutrition literacy, more transparent food labelling regulation, and evidence-based nutritional recommendations. Finally, replacing refined sugar with turbinado sugar does not dismiss health risks but rather decreasing total added sugar consumption and promoting whole-food-based dieting as the most ideal method of public health promotion.

Biosynthesized silver nanoparticles from Andrographis echioides leaf extract with promising antibacterial, antibiofilm, and cytotoxic properties

Sivagami Varatharajan, Uma Chinnaiyan, Sowndarya Sivaprakasam, Ethazhya Uthayasuryan, Mathan Chandrasekaran, Abinaya Vijayan, Sivasubramaniyan Manickam, Sivagurunathan Paramasivam

APJMBB 34(2): 182-194

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.16

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Silver nanoparticles (AgNPs) have gained a prominence among biological nanotechnology due to their ease of synthesis, stability, biocompatibility, and broad-spectrum activity. In this study, AgNPs were fabricated using a green synthesis approach based on Andrographis echioides leaf extract. The nanoparticles were studied using UV-Vis spectroscopy, FT-IR, Scanning Electron Microscopy (SEM), and zeta potential analysis. The UV-Vis spectra confirmed nanoparticle formation, with a distinct peak at 562 nm. FT-IR analysis revealed the functional groups that are involved in AgNP reduction and stabilization. SEM analysis revealed that the nanoparticles were predominantly spherical, with an average particle size of 149.0 nm. The zeta potential was recorded at -15.9 mV, indicating moderate colloidal stability. The biological potential of AgNPs was assessed using antibacterial, antibiofilm, and cytotoxic assays. The nanoparticles demonstrated effective antibacterial activity against four bacterial strains, with Bacillus subtilis and Pseudomonas aeruginosa being the most susceptible. The Congo red agar method demonstrated that 100 µg/mL of AgNPs effectively inhibited biofilm formation by P. aeruginosa, B. subtilis, S. aureus, and E. coli. MTT and LDH assays on the human fibroblast (MCCOY) cell line revealed strong anticancer activity. The MTT assay yielded an IC₅₀ value of 52.71 µg/mL, indicating dose-dependent cytotoxicity. The LDH assay supported membrane integrity disruption. These findings demonstrate the multifunctional applications of biosynthesized AgNPs, emphasizing their potential as antibacterial, antibiofilm, and anticancer agents in future biomedical and pharmaceutical research.

Halal authentication of African catfish (Clarias gariepinus) in Malaysian wet markets using porcine DNA detection 

Siti Nuradilah Mohd Rashid, Mohd Mahyeddin Mohd Salleh, Mohd Zuhaifah Mohamed Jamil, Wan Kamilah Wan Ibrahim, Nazariyah Yahaya

APJMBB 34(2): 195-205

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.17

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The African catfish (Clarias gariepinus) is a widely preferred species utilized in Malaysian aquaculture due to its adaptability and resilience to various environmental conditions. However, economic strains have led some fish breeders to use lower-quality feed that raises concerns about the halal integrity of the fish. This study aimed to investigate the presence of porcine DNA in African catfish sold at local markets, ensuring they meet halal standards for Muslim consumers. In this study, we extracted DNA from different tissues, flesh, stomach, and fins, to determine which offered the highest concentration. Notably, the fin showed the highest yield at 241.33 ng/µL. We used real-time polymerase chain reaction (RT-PCR) for porcine DNA detection because of its specificity and sensitivity. Out of 18 catfish samples tested, five showed positive for porcine DNA, indicating possible contamination linked to pig-derived feed. Interestingly, these samples had distinct characteristics, such as a yellowish hue and an unusual odor. This research underscores the need for strict DNA extraction protocols to ensure quality and compliance with halal standards, highlighting the importance of regulatory measures in aquaculture to protect Muslim consumers and support halal food markets globally.

From human pathogen to protector: Recent advances in the immunogenic potential of pneumococcal-derived extracellular membrane vesicles (EMVs)

Nurul Anis Johari, Nurul Fathiyah Zaipul Anuar, Fadzilah Mohd Nor, Mohd Nasir Mohd Desa, Eng Hwa Wong, Vanitha Mariappan, Kumutha Malar Vellasamy, Norhayati Liaqat Ali Khan, Jamal Hussaini, Navindra Kumari P

APJMBB 34(2): 206-215

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.18

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Streptococcus pneumoniae is a significant human pathogen globally, associated with various pneumococcal infections, implicated in conditions such as pneumonia, meningitis, otitis media and sepsis. The pathogen employs numerous virulence factors, such as capsular polysaccaharide (CPS), choline binding protein (CBP), lipoproteins and pneumolysin, to engage with and evade host immune response. Recent studies have emphasised the significance of extracellular membrane vesicles (EMVs) in pathogenesis and immunity, making them potential targets for innovative treatment and vaccination approaches. This review provides a comprehensive overview of EMVs derived from S. pneumoniae and delves into their immunogenic potential as a promising novel vaccine candidate against pneumococcal infections. This review describes the mechanisms of EMVs biogenesis, their various immunogenic properties and their capability to influence host immune responses. This review synthesises findings of the past 10 years (2014-2024) regarding S. pneumoniae EMVs, emphasising their biogenesis, composition, immunogenic properties and roles in host-pathogen interactions. Besides, this review also elaborates on EMV-based vaccines and the challenges associated with their advancement. The research underscores the diverse functions of EMVs derived from S. pneumoniae in pathogenesis, encompassing biofilm formation, immunological evasion and host cell damage. EMVs exhibit strong immunogenic characteristics capable of provoking both innate and adaptive immune responses. Pre-clinical studies have indicated EMVs as a broad-spectrum vaccination candidate, overcoming the constraints of existing serotype-specific vaccines. Nonetheless, challenges such as variations in EMVs’ composition, safety issues and production scalability persist as considerable barriers. EMVs are promising strategies in the global control of pneumococcal infections because of their ability to elicit cross-serotype immunity, yet further study is required to solve safety and manufacturing issues.

Integrative natural approaches for ischemic stroke: Evidence from plants, honey, marine bioproducts, and mushrooms

Nur Athirah Azlan, Misya Afiqah Noor Tuah, Nik Nasihah Nik Ramli, Norshafarina Shari, Muhammad Danial Che Ramli

APJMBB 34(2): 216-240

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.19

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Ischemic stroke remains a leading cause of long-term neurological disability, with limited therapeutic options for neuronal recovery. This review aims to critically evaluate natural products with neuroprotective potential against ischemic stroke including plants and herbs, bio-marine resources and mushrooms in stroke control. This review synthesizes evidence from peer-reviewed preclinical studies retrieved from databases including PubMed, Scopus, and Web of Science concentrating on preclinical experimental evidence. These natural products exhibit neuroprotective properties mainly through antioxidant, anti-inflammatory, and anti-apoptotic actions, as well as by influencing neurogenesis and synaptic plasticity.  Natural products exert neuroprotective effects primarily through modulation of oxidative stress, neuroinflammation, apoptotic pathways, and neurogenesis. Overall, the growing body of evidence underscores the therapeutic potential of natural products as complementary treatments for ischemic stroke, though additional translational and clinical research is necessary to confirm their efficacy and safety.

Study of TNIP1, CSK, and BANK1 gene expression in systematic lupus erythematosus in Iraqi children

Noor Mahmood Majeed, Widad Abed Jawad, Anwar Noori Ayoob

APJMBB 34(2): 241-246

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.20

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SLE, or systemic lupus erythematosus in children, is a very severe long-term autoimmune disease where the body's own tissues are attacked by the immune system, and can lead to lupus nephritis, cardiovascular problems, and neurological complications. The study aims to determine the changes in gene expression of TNIP1, CSK and BANK1 in children with SLE and to compare them with healthy controls, thereby contributing to the understanding of lupus pathogenesis. The total number of samples is 75 serum samples, of which 50 were collected from children attending hospitals who had clinical signs of lupus. In addition, 25 samples were collected from healthy children. The samples were kept in TRIzol to preserve the mRNA. The mRNA was transcribed into cDNA for the determination of gene expression. The results showed that the distribution of SLE in females (66%) was higher than the distribution of SLE in males (34%). The gene expression of TNIP1 and CSK was significantly decreased in SLE children compared to healthy children. At the same time, our work demonstrated that gene expression of the BANK1 gene was significant increased compared to that of healthy children. In conclusion, the gene expression of TNIP1, CSK, and BANK1 differs in SLE children compared to healthy children, which may assist researchers and clinicians in using these genes as potential biomarkers of SLE detection. The findings are useful for understanding the molecular aspects of the disease and may in the future contribute to improved diagnostic and treatment approaches targeting these genes.

Molecular detection of rickettsial diseases in a Malaysian hospital

Saadiah Abd Malek, Muhamad Yazli Yuhana, Nor Akmal Binti Mokhtar, Noor Hanisa Harun, Hazrul Nizam Hisham Hasmi, Siti Farah Alwani Mohd Nawi, Siti Norbahiyah Awad, Seok Mui Wang

APJMBB 34(2): 247-251

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.21

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Rickettsial infections are neglected and underdiagnosed in Malaysia. The epidemiological data and the exact burden of rickettsial infection in Malaysia are limited. The conventional serology method is usually used to diagnose the infection. However, the method is laborious and requires reference serum and antigens. Moreover, it can only detect a limited number of rickettsia species and strains. Hence, molecular genotyping is the method of choice for studying the genetic diversity of rickettsia as it can detect the causative agents at the genomic level. This study aimed to identify and characterize rickettsia species among adults presented to Teluk Intan Hospital, Perak, Malaysia, by polymerase chain reaction and DNA sequencing. A total of 141 Malaysian adult blood samples that fulfil the inclusion and exclusion criteria were recruited. These samples were subjected to PCR for scrub typhus (Orientia tsutsugamushi) by detecting the presence of 56-kDa type-specific antigen (TSA) gene and other rickettsiae by detecting the presence of 17-kDa. Only one sample was detected positive for scrub typhus, and sequence-verified to have a closet match with Orientia tustsugamushi strain UT302, which was originally isolated from Thailand. The number of positive cases was lower than anticipated, this was most likely due to the consequences of the global COVID-19 pandemic since December 2019, which coincide with the period of sample recruitment. The restriction control movement implemented by the Malaysian government and the Hospital being gazetted as a COVID-19 hospital has greatly affected the patient recruitment for this study.

Antimicrobial effect of Psidium guajava leaf against bacteria causing armpit malodors 

Hassanain Issam Al-Talib, Siti Zulaika Zulkeplee, Azlin Sham Ramly

APJMBB 34(2): 252-258

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.2.22

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Human body odor occurs when the microflora of the skin degrades the odorless fatty acid produced by the apocrine glands into volatile odor molecules. This study aimed to evaluate the antibacterial activity of methanolic leaf extract of Psidium guajava against odor-producing axillary microflora, including Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis, and Corynebacterium xerosis, using the agar well diffusion method for susceptibility testing and the broth microdilution method to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The study showed that all tested bacteria were sensitive to the leaf extract. S. epidermidis exhibited a higher MIC 0.68 mg/mL compared to S. aureus 0.22 mg/mL, whereas the remaining bacteria demonstrated an MIC of 2.05 mg/mL. The MBC for S. aureus was 6.17 mg/mL, while the other bacterial species showed a uniform MBC of 2.05 mg/mL. These findings suggest that guava leaf extract may serve as a potential alternative antibacterial agent for incorporation into cosmetic products, potentially replacing synthetic compounds.

Volume 34(3); 2026

Serum vitamin D levels and their association with glycemic control and insulin resistance in type 2 diabetes

Enny Probosari, Hertanto Wahyu Subagio, Heri Nugroho, Banundari Rachmawati, Siti F. Muis, Etisa A. Murbawani, Tri I. Winarni

APJMBB 34(3): 1-10

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.01

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Numerous studies have demonstrated the role of vitamin D in patients with type-2 diabetes mellitus (T2DM), but evidence regarding its association with glycemic control and insulin resistance among T2DM patients in Indonesia is still limited. This study investigated the association between serum 25-hydroxyvitamin-D [25(OH)D] levels, glycemic control, and insulin resistance among patients with T2DM. A cross-sectional study was conducted among 129 T2DM patients aged 30–80 years in Semarang, Indonesia. Serum 25(OH)D and insulin levels were measured using ELISA, fasting plasma glucose (FPG) with the GOD-PAP method, glycated hemoglobin (HbA1c) using enzyme immunoassay, and insulin resistance assessed with HOMA-IR. Correlation tests and multivariate linear regression analyses were performed, adjusting for age, body mass index, and sex. Vitamin D deficiency was found in 48.8% of subjects, predominantly in females. Serum 25(OH)D levels were inversely correlated with HbA1c (r = –0.257, p = 0.003) and FPG (r = –0.173, p = 0.050). After adjustment, significant negative associations remained for HbA1c (B = –0.020, p = 0.009) and FPG (B = –0.942, p = 0.007). Sex-stratified analyses showed that these inverse associations were significant only in females, while in males, 25(OH)D was positively associated with fasting insulin. No significant relationship was found between 25(OH)D and HOMA-IR. These findings suggest that serum vitamin D levels are associated with glycemic control in patients with T2DM, with potential sex-specific differences, underscoring the need to consider vitamin D status in diabetes management strategies.

Molybdenum disulfide-enhanced bacterial cellulose through static fermentation: Optimisation and structure–property relationships

Nur Farhanul Iman Ahmad Fozi, Farzana Muhammad Ramzan, Nuratika Hasanuddin, Nurul Syafikah Putri Mohd Jeffree, Muhammad Iffat Hanif Mohd Rudin, Azila Adnan

APJMBB 34(3): 11-20

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.02

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Bacterial cellulose (BC) is a high purity nanofibrillar biopolymer with promising applications in textiles, packaging, and biomedicine. Here we report a fermentation-integrated strategy to modulate BC production and structure by introducing molybdenum disulfide (MoS₂) nanosheets during SCOBY (symbiotic culture of bacteria and yeast) cultivation. MoS₂ was added at concentrations of 0.2–0.6% (w/v) and compared against an untreated control under static conditions (30°C, pH 5.0, 14 days). Production metrics, FTIR spectroscopy, and X-ray diffraction (XRD) were employed for characterisation. At 0.3% MoS₂, volumetric productivity increased from 4.2 to 11.3 g·L^-1·day^-1 (≈169% increase, p < 0.05). FTIR analysis revealed a band at 1033 cm^-1, indicating partial MoS₂ oxidation (S=O/Mo–O vibrations) and interfacial bonding with cellulose. At concentrations ≥0.4%, nanoparticle aggregation reduced productivity and crystallinity. This work demonstrates a bio-fabrication route to tune BC structure-property profiles via inorganic nanosheets and elucidates dose-dependent mechanisms balancing microbial activity with filler dispersion.

Mutation-resilient RBD binding and host protection: Piperine for pediatric COVID-19

Shu-wei Beh, Yu Heng Liew, Gideon Khoo, Lyn R Griffiths, Loo Keat Wei

APJMBB 34(3): 21-32

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.03

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COVID-19 presents an escalating threat in children, manifesting as persistent respiratory, neurological, and multisystem complications, alongside elevated risks of myocarditis, thromboembolism, and autoimmune dysregulation. Emerging Omicron subvariants, including BA.2, BA.2.86, and JN.1, exhibit pronounced immune evasion, undermining current therapeutics. Pediatric populations are particularly vulnerable due to immature immune and metabolic systems, and the safety profile of nirmatrelvir in this group remains inadequately characterized. Here, we identify piperine, alone or in combination with curcumin, as a dual-function, pediatric-compatible phytopharmaceutical. Molecular docking across B.1.1.529, BA.2, and a homology-modeled JN.1 receptor binding domain (RBD) demonstrated that piperine forms low-energy, structurally stable complexes (ΔG = – 8.46 kcal/mol), in contrast to nirmatrelvir (ΔG = – 7.20 kcal/mol), and consistently engages mutationally constrained RBM and core residues. Piperine retains robust binding even though structural analyses reveal that coordinated mutations (S371F, T376A, V445H, G446S, L452W, E484K, F486P) remodeled the β-sheets and α-helices of the RBD, which may affect ACE2 and antibody interactions. Complementary ADME and drug-likeness assessments further support its oral bioavailability and dietary suitability. Collectively, these findings position piperine, with or without curcumin, as a dual-action therapeutic, i.e., combining mechanistic antiviral activity through mutation-resilient RBD engagement with systemic protective effects. This strategy highlights its potential as a safe, diet-compatible adjunct to mitigate COVID-19 sequelae in children, offering a translationally relevant approach for variant-adapted pediatric care.

Upregulation of NFκB2 transcription factor gene towards human norovirus virus-like particle replication in microfold cells three-dimensional co-culture model

Emilly Sion, Sharaniza Ab-Rahim, Mudiana Muhamad

APJMBB 34(3): 33-50

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.04

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Human norovirus (HuNoV) is the leading cause of non-bacterial acute gastroenteritis, posing substantial socioeconomic and health burdens. A comprehensive understanding of HuNoV pathobiology remains limited due to the virus’s non-cultivatable nature in conventional in vitro systems. Toll-like receptors (TLRs) participate in the host cell invasion of enteric viruses and initiate immune responses through specific transcription factors; however, the factors and mechanisms relevant to HuNoV remain unclear. Thus, this study aims to investigate the mechanistic association between the TLRs and corresponding transcription factors. Due to limited availability of HuNoV isolates, this study resorted to HuNoV-VLP, constructed by seamless cloning technique, inoculated into the M cells 3D Alcart^©, a 3-dimensional culture system established previously. Morphological changes of the 3D model attributable to HuNoV action was validated using electron microscopy (EM) and immunofluorescence. Real Time qPCR was performed to measure the expression of the transcription factor genes. EM and immunofluorescence results showed that the M cells 3D Alcart^© model supported HuNoV-VLP uptake, intracellular localization and processing, indicated by presence of white-stringy structure and virus particles in the cytoplasm, with increased expression of M cells marker, UEA-1. Gene expression analysis revealed a significant upregulation in the expression of NF-κB2 at 24 hours post-inoculation, with no significant changes observed for NF-κB1, IRF3, and IRF7. In conclusion, HuNoV-VLP replication within the M-cell 3D Alcart^© co-culture model appears to selectively activate the noncanonical NF-κB2 pathway without broadly engaging other antiviral transcription factors.

A multi-epitope mRNA vaccine candidate against Trichinella spiralis: An integrated immunoinformatics and  computational approach

Roohollah Fattahi

APJMBB 34(3): 51-61

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.05

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Trichinella spiralis is a parasitic nematode causing trichinellosis, a significant zoonotic disease, and the development of an effective vaccine remains a crucial goal. This study employed a comprehensive computational approach to design a multi‑epitope mRNA vaccine against T. spiralis. From 49,084 T. spiralis proteins retrieved from NCBI, secreted and membrane proteins were identified using SignalP 6.0, DeepLoc 2.1, and TMHMM. Antigenic proteins were shortlisted with VaxiJen. B‑cell, cytotoxic T‑lymphocyte (CTL), and helper T‑lymphocyte (HTL) epitopes were predicted using ABCpred and IEDB servers. The vaccine construct was designed with an adjuvant, selected epitopes, and necessary mRNA elements. Its antigenicity, allergenicity, and physicochemical properties were evaluated. Codon optimization, secondary and tertiary structure prediction, molecular docking with Toll‑like receptors (TLRs), molecular dynamics simulations, and immune response profiling were performed. From 2,991 secreted proteins, nine highly antigenic proteins were selected, yielding 36 epitopes (18 B‑cell, 9 CTL, 9 HTL), all confirmed as antigenic, non‑allergenic, and non‑toxic. The designed vaccine construct is antigenic (VaxiJen score 0.84), non‑allergenic, non‑toxic, and stable. Molecular docking revealed strong binding affinities with TLR2, TLR3, and TLR4; normal mode analysis confirmed complex stability; immune simulation predicted robust activation of cellular and humoral immune responses, including elevated cytokines and immunoglobulins. The proposed multi‑epitope mRNA vaccine candidate demonstrates high immunogenic potential and stability against T. spiralis, warranting further experimental validation.

Comparative assessment of growth and antioxidative defense mechanisms in twelve Centella asiatica callus lines 

Norhayati binti Yusuf, Aziz Ahmad

APJMBB 34(3): 62-77

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.06

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Antioxidants in fruits and vegetables play a critical role in protecting cells against oxidative stress. Centella asiatica (pegaga) is well known for its high antioxidant and medicinal properties. This study examined the enzymatic, non-enzymatic, and free radical scavenging activities of C. asiatica callus cultures derived from twelve accessions (CA01-CA12). Considerable variability was observed among accessions, reflecting strong genotype-dependent regulation of oxidative defense mechanisms. CA08 accumulated the highest ascorbic acid and α-tocopherol, whereas CA12 contained the most carotenoids. Glutathione content varied markedly among C. asiatica accessions, with CA03, CA05, and CA08 exhibiting the highest glutathione content. Enzyme assays revealed distinct antioxidant capacities: CA09 exhibited exceptionally high superoxide dismutase (SOD) and catalase (CAT) activities, indicating strong detoxification of superoxide and hydrogen peroxide, whereas CA01 and CA11 showed elevated glutathione reductase (GR) and ascorbate peroxidase (APX) activities, supporting efficient recycling of the glutathione-ascorbate system. Total antioxidant capacity, free radical scavenging assays (DPPH, superoxide, and hydroxyl radicals) confirmed substantial antioxidant potential in most callus lines, with several showing activities comparable to the control. The wide range of hydroxyl radical scavenging further emphasized accession-specific defense strategies. Overall, C. asiatica callus cultures represent a promising in vitro source of antioxidant compounds.

Modulating Escherichia coli cell attachment via metabolic flux redistribution of the PTA-acetyl phosphate pathway under phosphate addition and oxygenation levels 

Nurul Febriani Putri, Prayoga Suryadarma, Fitri Handayani, Fitriagustiani, Muh. Chaeril Ikramullah, Pasmawati

APJMBB 34(3): 78-87

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.07

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The effect of phosphate addition in cultivation media on optimizing Escherichia coli cell attachment was investigated. Cultivation media were prepared using minimal media without phosphate, followed by adding various phosphate concentrations (0, 1.33, 3.32, 6.64, and 9.97 g/L). Cell attachment was evaluated by measuring the index of colonizing cells (Ics) in a PVC 96-well plate, along with dry cell weight (DCW), phosphate consumption, and carbon flux. The influence of oxygenation was assessed at two phosphate concentrations (0 and 9.97 g/L) with different agitation levels. Results showed that 9.97 g/L phosphate significantly enhanced cell attachment, increasing the Ics to approximately 0.698. This was accompanied by a decrease in TCA cycle activity, indicated by a reduced DCW of 0.049 g/L, suggesting phosphate was used via the PTA pathway to synthesize acetyl phosphate. Increasing oxygenation up to 200 rpm further improved attachment, raising Ics to 0.848 and increasing DCW to 0.109 g/L due to enhanced cell mass synthesis. In conclusion, we suggest that phosphate addition likely modulates cell attachment, potentially via the PTA-Acetyl phosphate pathway, while higher oxygenation supported attachment by stimulating cell growth.

Polyphenol-rich trihoney modulates hematological and lipid profiles in hyperlipidemic rabbits 

Ainin Azwani Abdul Rafa, Nor Hafizah Zakaria, Muhammad Ibrahim, Mohd Nur Nasyriq Anuar, Fadzilah Adibah Abdul Majid, Radiah Abdul Ghani

APJMBB 34(3): 88-100

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.08

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This study investigates the effects of Trihoney, a blend of honey from Trigona sp., Mellifera sp., and Dorsata sp., on hematological and lipid profiles in hyperlipidemic New Zealand male white rabbits. Polyphenols in Trihoney were identified and quantified using ultra-high-performance liquid chromatography (UHPLC). Thirty rabbits were divided into six groups and fed different diets for 12 weeks. Four groups received a 1% high-cholesterol diet (HCD): one served as the HCD control, one received atorvastatin, and two were treated with Trihoney at 0.3 g/kg body weight/day (T1) or 0.6 g/kg body weight/day (T2). The remaining groups were fed a commercial pellet diet, with one also receiving 0.6 g/kg body weight/day of Trihoney. Blood samples were collected at baseline and after 12 weeks. Cinnamic acid and hydroxybenzoic acid were the most abundant polyphenols, with concentrations of 4340 ng/mL and 2850 ng/mL, respectively. All HCD-fed rabbits developed hyperlipidemia. Trihoney supplementation improved red blood cell count, hemoglobin, and hematocrit. The higher dose (T2) also normalized elevated white blood cells, lymphocytes, monocytes, and platelets. Total cholesterol (TC) increased across treatment groups, but the HCD+T1 group showed the smallest increase. Trihoney at 0.3 g/kg body weight/day reduced triglycerides (TG) by 30% (p < 0.05). Both LDL and HDL cholesterol levels rose in Trihoney- and atorvastatin-treated rabbits, though increases were less pronounced compared to the untreated HCD group. Trihoney shows potential as a natural dietary intervention to improve lipid and hematological parameters in hyperlipidemic conditions, suggesting a protective role against cardiovascular complications.

Comparative study on cytotoxic activity of Acanthamoeba clinical and environmental isolates lysate on MCF-7 cells 

Nuraini Nawi, Fadzilah Adibah Abdul Majid, Nor Hafizah Zakaria

APJMBB 34(3): 101-114

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.09

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Breast cancer is the most diagnosed cancer among women worldwide, driving the search for more effective treatments and the discovery of novel therapeutic agents from diverse biological sources, including protozoa such as Acanthamoeba. Our preliminary findings demonstrated that Acanthamoeba lysates disrupt cancer cell signaling pathways and induce apoptosis in these cells. Building on these results, the present study aims to comprehensively compare three isolates, Acanthamoeba castellanii (IMR), Acanthamoeba sp. (HKL), and Acanthamoeba sp. (SW) with respect to their growth characteristics and protein profiles, and to correlate these features with their cytotoxic activities. The study showed that day five was the optimal time for trophozoite collection, which exhibited the highest area fraction (34.93%) before entering the stationary phase. This aspect, examined for the first time, contributes to the standardization of lysate preparation. Protein profiling using SDS-PAGE and cluster analysis revealed significant strain-specific differences. The IMR and SW isolates appeared closely related, while HKL displayed distinct banding patterns. These variations offer new insights into molecular diversity that may affect anti-cancer potency. Cytotoxicity assays conducted on MCF-7 breast cancer cells showed a significant reduction in cell viability following treatment with the lysates, with the SW isolate demonstrating the most robust effect (IC₅₀ = 15.09 ± 1.12 μg/ml), outperforming both IMR and HKL. This increased effectiveness emphasizes the importance of selecting the appropriate strain for identifying bioactive compounds. Apoptosis was confirmed as the primary mechanism of cell death, as indicated by DNA fragmentation, morphological changes, and AOPI staining. These findings reinforce prior observations and demonstrate that apoptotic activity is correlated with distinct protein signatures across the different isolates. In conclusion, this study extends earlier work by establishing optimal conditions for trophozoite harvesting, revealing protein-based strain variations, and identifying the SW isolate as the most promising candidate for apoptosis induction in anti-cancer research.

Targeting ERK activity in allergic inflammation: Biological roles and natural compound-based inhibition

Lisbeth Su Yee Kong, Salah A. Alshehade, Ravishankar Ram Mani, Malarvili Selvaraja, Sasikala Chinnappan, Ming Tatt Lee, Chau Ling Tham, Audrey Siew Foong Kow

APJMBB 34(3): 115-127

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.10

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Allergy is a prevalent immunological disorder that affects individuals of various age groups, imposing continuous economic and health burdens globally. Extracellular signal-regulated kinases 1 and 2 (ERK1/2) serve as critical downstream effectors in the Ras-Raf-MEK-ERK cascade, orchestrating allergic inflammation through multiple cell-specific mechanisms. Following IgE-FcεRI crosslinking on mast cells and basophils, ERK1/2 phosphorylation by MEK1/2 triggers immediate degranulation and release of preformed mediators including histamine, β-hexosaminidase, and proteases. Upon activation, phosphorylated ERK1/2 translocate to the nucleus where they phosphorylate transcription factors including Elk-1, c-Fos, c-Jun, and NFAT, driving expression of pro-inflammatory genes encoding TNF-α, IL-4, IL-5, IL-6, and IL-13. In T lymphocytes, ERK1/2 signaling governs naïve CD4+ cell differentiation into Th2 effector cells, thereby promoting IgE synthesis and perpetuating allergic sensitization. In eosinophils, IL-5-induced ERK1/2 activation enhances survival, migration, and inflammatory mediator release. Given ERK1/2’s central position in allergic pathophysiology, selective inhibition represents a rational therapeutic strategy. Plant-derived phenolic compounds, including flavonoids (quercetin, luteolin, baicalein), curcuminoids (curcumin), and phenolic acids (rosmarinic acid), have demonstrated capacity to suppress ERK1/2 phosphorylation, thereby attenuating mast cell degranulation, reducing cytokine production, and modulating Th1/Th2 balance. This review examines ERK1/2’s molecular mechanisms in allergic inflammation and evaluates natural phenolic compounds as ERK-targeted therapeutic interventions for allergic diseases.

Indian costus-mediated synthesis of silver nanoparticles: Characterization and anti-staphylococcal activity with focus on DNA damage

Sabaa Ali Mohammed Al. Fadal, Rafeef Amer Abdul-Jabar, Enas A. Bady

APJMBB 34(3): 128-140

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.11

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Preparation of nanoparticles using plant extracts plays an important role in nanotechnology due to its low cost and eco-friendliness. In this study, silver nanoparticles were prepared using an aqueous extract of Indian costus, and their destructive effect on Staphylococcus aureus DNA isolated from diabetic foot ulcers was evaluated. An aqueous extract of Indian costus was prepared using the reflux method, and its flavonoid and phenolic acid contents were assessed. Silver nanoparticles were synthesized using a 3% aqueous extract of Indian costus and characterized after 24 hours using UV-Vis spectrophotometry, Zetasizer analysis, Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). Biological activities were evaluated through DPPH antioxidant, antibacterial, and DNA cleavage assays against Staphylococcus aureus DNA isolated from diabetic foot ulcers. The synthesized silver nanoparticles were characterized by UV-Vis spectroscopy, while SEM analysis revealed a spherical morphology and FTIR analysis confirmed the presence of silver oxide (AgO), indicating successful nanoparticle formation. The produced silver nanoparticles demonstrated antibacterial activity against Escherichia coli, Staphylococcus aureus, and Candida albicans at concentrations of 1, 0.5, and 0.25 mg/mL. In addition, AgNPs exhibited DNA cleavage activity against Staphylococcus aureus isolated from diabetic foot ulcers at the same concentrations. The nanoparticles demonstrated antioxidant and antibacterial activities, as well as a strong effect on Staphylococcus aureus DNA isolated from diabetic foot ulcers. These activities were associated with a moderate hemolytic effect, particularly at higher concentrations.

Mangrove root fungal endophytes as a source of biomass degrading enzymes

Heri Ariadi, Tri Yusufi Mardiana, Linayati Linayati, Benny Diah Madusari, M. Bahrus Syakirin

APJMBB 34(3): 141-148

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.12

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Fungal endophytes isolated from mangrove roots demonstrated strong halotolerant characteristics, as indicated by their ability to maintain growth under a wide range of NaCl concentrations (0, 1, 2, 3, and 4%). Although several fungal isolates exhibited a gradual reduction in growth performance as salinity increased, Lasiodiplodia theobromae and Pseudopestalotiopsis theae showed stable growth responses across all tested salinity levels, suggesting their high physiological adaptation to saline environments. Interestingly, Phomopsis sp. 1 exhibited enhanced growth under salt stress conditions, indicating that certain mangrove-associated endophytes may possess specific adaptive mechanisms that allow them to utilize or tolerate elevated salt concentrations. In addition to their halotolerance capability, several mangrove root endophytes demonstrated functional enzymatic activities by producing salt-induced or salt-tolerant cellulase and β-glucosidase enzymes. These fungi were also capable of producing laccase enzymes that remained active under saline conditions. The ability to maintain extracellular enzyme activity under high salinity conditions highlights the ecological importance of these fungi in mangrove ecosystems, particularly in environments characterized by fluctuating salinity and prolonged inundation. The presence of salt-tolerant lignocellulolytic enzymes suggests that mangrove root endophytes play an essential role as biological decomposers, facilitating the breakdown of complex organic materials such as cellulose and lignin derived from plant residues. Consequently, these fungi contribute significantly to nutrient mineralization and biogeochemical cycling within submerged mangrove ecosystems. Their adaptation to saline habitats also indicates potential applications as functional microbial resources for biotechnology, particularly in the development of environmentally sustainable biomass degradation processes.

In vitro antifungal activity of ethanolic fingerroot (Boesenbergia rotunda) extract against the phytopathogen Paramarasmius palmivorus

Takdanai Sakulmettathum, Onanong Pringsulaka, Tripatchara Atiratana, Sutheewan Binchai, Achariya Rangsiruji

APJMBB 34(3): 149-159

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.13

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Crude extracts from four Zingiberaceae species were investigated for their antifungal activity against Paramarasmius palmivorus, a phytopathogenic fungus affecting a wide range of economically important crops. Ethanolic extracts of Alpinia galanga, Boesenbergia rotunda, Curcuma longa, and Zingiber officinale were prepared by maceration and tested in vitro using the poisoned food technique. Ten-fold serial dilutions of each extract (100,000–10 ppm) produced concentration-dependent inhibition of mycelial growth. Among the four species, B. rotunda exhibited the greatest antifungal activity, completely inhibiting mycelial growth at 100,000 ppm and resulting in significantly lower mycelial growth than the other extracts at the higher concentrations tested. The minimum inhibitory concentration of the B. rotunda extract was 25,000 ppm. In comparison, C. longa and Z. officinale exhibited moderate antifungal activity, whereas A. galanga showed the weakest inhibitory effect. Although B. rotunda was the most effective crude extract evaluated, the relatively high concentration required for complete inhibition indicates that further optimisation of extraction methods and formulations, together with validation under greenhouse and field conditions, is required before practical agricultural application can be considered.

Determination of urinary mitragynine concentration and severity of kratom dependence among people with kratom dependence in Malaysia

Jia-Qian Ang, Chia-Yee Lok, Sue-Hay Chan, Nik Hisamuddin Nik Ab. Rahman, Mohd Azar Mohd Yasin, Lay-Harn Gam

APJMBB 34(3): 160-170

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.14

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Kratom is used traditionally for its medicinal properties in Southeast Asia. The main alkaloid content of kratom leaves is mitragynine, and it binds to opioid receptors. Therefore, kratom exerts opioid-like effects and carries a risk of addiction if used for a long period of time. In this study, we aimed to evaluate the urinary mitragynine of people with kratom dependence and relate it to habitual use of kratom. The severity of kratom dependence among respondents is assessed using the Kratom Dependence Score method. The concentration of mitragynine in urine was analyzed using LC/MS/MS. Significant variation in urinary mitragynine concentration was observed, ranging from the lowest of 58 ng/mL to the highest of 18,000 ng/mL. Urinary mitragynine concentration showed no significant association with the severity of kratom dependence. While Kratom Dependence Score analysis revealed a positive association between the severity and duration of kratom use (p=0.024). Many (36%) of people with kratom dependence confessed to recreational kratom use in their twenties. Urinary concentration of mitragynine does not correlate with severity (H=1.267, df=2, p=0.531) and long-term kratom dependency (H=0.860, df=3, p=0.835)

Bacteriophage therapy as an alternative to antibiotics for controlling necrotic enteritis in poultry: A comprehensive review

Muhammad Zulfaiz Zuikarnain, Mohd Asrore Mohd Shaufi, Nurul Aliesa Azzan, Suet Lin Chia, Hasliza Abu Hassim, Mohd Termizi Yusof, Shuhaimi Mustafa

APJMBB 34(3): 171-181

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3.15

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Necrotic enteritis (NE), caused by Clostridium perfringens, remains a significant challenge in poultry farming, causing reduced productivity and economic losses globally. While antibiotics have been a mainstay in NE control, their use is increasingly restricted due to rising antimicrobial resistance and consumer demand for antibiotic-free products. This has driven the search for effective alternatives, with phage therapy emerging as a promising solution. This review examines phage therapy's potential in combating C. perfringens-induced NE, highlighting its specificity, adaptability, and minimal impact on beneficial gut microbiota. Key findings reveal that multivalent phage cocktails and higher phage concentrations yield better outcomes than single-phage approaches, while encapsulation techniques enhance phage stability and efficacy during gastrointestinal transit. Optimal delivery through feed or water and pre-emptive administration during critical growth phases significantly improve therapeutic success. However, challenges such as phage stability, scalable production, and regulatory hurdles must be addressed to facilitate widespread adoption. Advances in encapsulation and genetic engineering offer promising pathways to overcome these obstacles. By integrating phage therapy into poultry health management practices, it is possible to reduce reliance on antibiotics, improve animal welfare, and support sustainable poultry production. This review provides an in-depth analysis of current evidence and identifies future research priorities for advancing this innovative approach.

Volume 34 (3) Supplementary Issue 

Issue DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Suppl)

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This supplementary issue contain abstracts presented at International Conference on Molecular Biology and Biotechnology 2026 (ICMBB2026)

Volume 34(3) (Special Issue); 2026

Zinc oxide nanorods-polypyrrole nanocomposite as an effective photocatalyst for methylene blue dye degradation

Jingyou Ho, Zhan Peng Chu, Ai Ling Pang, Mei Lee Ooi, Mohsen Ahmadipour

APJMBB 34(3) (Special): 1-8

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Special).01

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Dye-contaminated effluents pose a serious threat to the environment, making it essential to address this issue to support environmental sustainability. The conventional methods for removing such pollutants include adsorption, flocculation/coagulation, ion exchange, sedimentation, reverse osmosis, membrane filtration, chemical precipitation, and various chemical or electrochemical treatments are found to be costly and time-consuming. In recent decades, heterogeneous photocatalysis has gained attention as a promising alternative. In this study, zinc oxide (ZnO) nanorods will be synthesized on kanthal wires using the direct heating (DH) method (at 90 W for 60 min), chosen for its simplicity, low energy consumption, and cost-effectiveness. The resulting ZnO nanorods will then be coated with polypyrrole (PPy) via electropolymerization with various coating durations (1 and 5 min) to create a nanocomposite material. The optical and surface morphology characteristics of the ZnO/PPy nanocomposites will be examined using UV-Visible Spectroscopy (UV-Vis) and Field Emission Scanning Electron Microscopy (FESEM). Their photocatalytic performance will be assessed by degrading 10 ppm and 20 ppm of methylene blue (MB) dye under UV light. The study found that the ZnO coated with PPy in 1 min duration was able to degrade approximately 89.72% of a 10 ppm MB solution within 3 hrs, demonstrating its potential as an effective and eco-friendly solution for water purification. This study utilized a hybrid method combining direct heating for rapid ZnO nanorods synthesis and the systematic optimization of PPy coating time via electropolymerization in investigating its influence on morphology, optical properties, and photocatalytic performance to fabricate the nanocomposites, which is a novel and practical approach.

Spent green tea residue–mediated green synthesis of zinc oxide nanoparticles (ZnO NPs): extract-dosage-controlled structural and biofunctional properties

Xiao-hui Chu, Sharon Chia Yen Lim, Ming Xiu Koh, Siew-Yoong Leong, Mohammod Aminuzzaman, XinGuang Fan, Ling Shing Wong, Lai-Hock Tey

APJMBB 34(3) (Special): 9-21

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Special).02

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Spent green tea residue represents an abundant post-consumption biomass that can be valorised as a sustainable resource for green nanomaterial synthesis. In this study, zinc oxide nanoparticles (ZnO NPs) were synthesised via a green route using spent green tea extract, with particular emphasis on extract dosage as a controlling parameter for structural and biofunctional properties. By varying the extract dosage from 0.02 to 0.05 g/mL, phase-pure wurtzite ZnO was obtained across all samples, while marked changes in crystallite size, defect characteristics, and optical response were observed. Increasing extract dosage progressively reduced crystallite size from approximately 52.5 to 30.9 nm and induced an apparent optical band-gap narrowing from 3.46 to 2.68 eV, possibly associated with defect- and surface-state contributions rather than any phase transformation. Biofunctional evaluation revealed a dosage-dependent enhancement in antioxidant activity, with DPPH radical scavenging efficiency increasing at higher extract dosages, consistent with strengthened surface-mediated redox interactions. In contrast, antibacterial performance exhibited a non-monotonic trend. ZnO NPs synthesised at an intermediate extract dosage (0.04 g/mL) demonstrated the strongest antibacterial activity against Gram-positive bacteria, producing inhibition zones of approximately 22–24 mm, whereas activity against Gram-negative bacteria remained limited under diffusion-based assay conditions. The reduced antibacterial efficacy at the highest extract dosage is attributed to increased aggregation and surface passivation effects, which suppress effective interfacial interactions. Overall, these findings establish extract dosage as a simple yet effective lever for tailoring the structure–property–bioactivity relationships of spent-tea-mediated ZnO NPs. This work supports sustainable nanomaterial development through waste valorisation, contributing to responsible production and biofunctional material innovation.

Comparative stability and oxidative potential of chromium (III) oxide (Cr2O3) nanoparticles (NPs), microcellulose microcomposites, and nanocellulose nanocomposites

Sharon Chia Yen Lim, Ming Xiu Koh, Ming Er Koh, Shi Yan Cheah, Xiao-Hui Chu, Ling Shing Wong, Lai-Hock Tey

APJMBB 34(3) (Special): 22-31

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Special).03

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The dispersion stability and oxidative behaviour of metal oxide nanomaterials are critical factors governing their performance, reproducibility, and safety in environmental and biological applications. Chromium (III) oxide (Cr₂O₃) nanoparticles (NPs), however, are inherently prone to aggregation and exhibit variable reactive oxygen species (ROS) generation in aqueous media, limiting their reliable deployment. In this study, the colloidal stability and oxidative potential of bare Cr₂O₃ NPs were systematically compared with cellulose-based composite systems, namely a Cr₂O₃–microcellulose microcomposite (MCP) and a Cr₂O₃–nanocellulose nanocomposite (NCP), to elucidate the role of matrix architecture in modulating nanoparticle behaviour. Chromium concentrations were quantified by inductively coupled plasma optical emission spectrometry (ICP–OES) to enable chromium-normalised comparisons across all formulations. Colloidal stability was evaluated using zeta potential and dynamic light scattering analyses, while dispersion state and re-aggregation behaviour were examined by transmission electron microscopy (TEM). ROS generation was assessed using a fluorescence-based hydroxyl radical (•OH) assay under controlled aqueous conditions. Despite exhibiting a high absolute zeta potential, Cr₂O₃ NPs displayed pronounced aggregation and inconsistent oxidative behaviour, highlighting the limitations of electrostatic stabilisation alone. In contrast, MCP and NCP demonstrated improved colloidal stability through cellulose-mediated steric effects, with nanocellulose providing the most effective long-term stabilisation via physical confinement. ROS assays revealed that MCP generated the highest and most sustained •OH levels, particularly under acidic conditions, likely due to partial surface exposure of Cr₂O₃ within the microcellulose matrix. Conversely, NCP exhibited minimal and rapidly diminishing ROS generation, consistent with effective surface shielding by the nanocellulose network. These findings demonstrate that composite architecture, rather than nanoparticle composition alone, governs the balance between dispersion stability and oxidative activity. Overall, this study provides fundamental insights into stability–reactivity relationships in Cr₂O₃-based systems and highlights matrix engineering as a rational strategy for designing safer and more predictable nanocomposites.

Green-synthesised metal oxide nanoparticles: Phytochemical mechanisms, structure–property relationships, and biofunctional applications

Xiao-hui Chu, Xinn Tze Chuah, Elice Zheng Rou Song, Sinouvassane Djearamane, Foo Win Yip, Mohammod Aminuzzaman, XinGuang Fan, Ling Shing Wong, Lai-Hock Tey

APJMBB 34(3) (Special): 32-48

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Special).04

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Green synthesis of metal oxide nanoparticles (NPs) using plant-derived extracts has emerged as a sustainable alternative to conventional fabrication routes while introducing distinctive surface chemistries that influence nanoparticle structure, redox behaviour, and biological interactions. Although numerous reviews have summarised green synthesis approaches and applications, the mechanistic links between phytochemical-mediated synthesis, nanoparticle physicochemical properties, and biofunctional performance remain insufficiently integrated. This review critically examines how plant secondary metabolites regulate nucleation, growth, stabilisation, and surface functionalisation of metal oxide NPs, with particular emphasis on zinc oxide (ZnO), copper oxide (CuO), and chromium oxide (Cr₂O₃) systems, which serve as representative models owing to their extensive mechanistic and biofunctional investigation. The relationships between particle size, crystallinity, defect density, surface charge, retained phytochemicals, and biological activities including antioxidant, antimicrobial, enzyme inhibitory, and cytotoxic responses are discussed within a unified structure–property–biofunction framework. Current evidence suggests that the biological performance of green-synthesised metal oxide NPs depends not only on their inorganic composition but also on phytochemical-derived surface chemistry, synthesis conditions, and exposure dose. Key challenges, including extract variability, insufficient surface characterisation, limited standardisation of biological evaluation, and translational safety considerations, are critically discussed together with future perspectives emphasising mechanism-driven synthesis, reproducibility, and harmonised safety assessment. Overall, green-synthesised metal oxide NPs should be regarded as chemically and biologically programmable nanomaterials that support the rational design of sustainable biofunctional materials for biomedical, biochemical, and environmental applications in alignment with the United Nations Sustainable Development Goals.

Morphology-dependent distribution of Cr₂O₃ nanoparticles on engineered graphene-based nanocarriers: Comparative SEM, FTIR and EDX characterisation

Ming Xiu Koh, Sharon Chia Yen Lim, Ming Er Koh, Shi-Yan Cheah, Xiao-hui Chu, Ling Shing Wong, Lai-Hock Tey

APJMBB 34(3) (Special): 49-58

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Special).05

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Graphene oxide (GO) and its derivatives are widely investigated as nanocarriers because their morphology and surface chemistry can be tailored for nanoparticle incorporation. Here, we examined how differences in graphene carrier morphology affect the distribution of chromium (III) oxide (Cr₂O₃) nanoparticles (NPs). Three graphene-based carriers, namely graphene oxide (GO), reduced graphene oxide (rGO), and spherical reduced graphene oxide (SrGO), were prepared using modified Hummers’ oxidation, chemical reduction, and aerosol spray-freezing approaches, respectively. Cr₂O₃ NPs were incorporated into each carrier under the same experimental conditions, and the resulting composites were characterised by Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). SEM revealed clear differences in Cr₂O₃ distribution among the three carriers. In particular, SrGO showed a more extensive and relatively homogeneous nanoparticle distribution across the observed regions, which coincided with its crumpled three-dimensional morphology and irregular surface features. Representative EDX measurements also showed differences in relative Cr abundance among GO–Cr₂O₃, rGO–Cr₂O₃,  and SrGO–Cr₂O₃. These EDX data were interpreted semi-quantitatively because replicate measurements were not available. Taken together, the findings show that graphene carrier morphology is associated with differences in the surface distribution of Cr₂O₃ NPs. The present results provide a basis for further quantitative studies of graphene–Cr₂O₃ nanocomposites and their potential use in catalysis, environmental remediation, and related applications.

Preliminary evaluation of oyster mushroom-supplemented yoghurt fermented with Lactobacillaceae strains: Lactic acid bacteria viability and inhibitory effect on Escherichia coli

Ana Mardiyaningsih, Widodo Widodo, Puji Astuti, Purwanto Purwanto

APJMBB 34(3) (Special): 59-67

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Special).06

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Members of the Lactobacillaceae family are well-established as beneficial microorganisms with significant functional value in the food and pharmaceutical industries. To ensure optimal health benefits, maintaining probiotic viability in the intestinal environment is crucial, a process that can be supported by prebiotic supplementation. The high cost of commercial prebiotics, like inulin and fructo-oligosaccharides, necessitates exploring affordable local alternatives. Oyster mushroom (Pleurotus ostreatus) is a potential source of non-starch polysaccharides, particularly β-glucans, known for their prebiotic and antibacterial properties. This study aimed to assess the impact of white oyster mushroom on the quality of yoghurt fermented with a mixed Lactobacillaceae starter culture, focusing on visual and physical characteristics, physicochemical characteristics (total protein and total solid) preliminary sensory acceptability, as well as its influence on LAB (lactic acid bacteria) growth and inhibitory effect against Escherichia coli (E. coli). An experimental design was employed, including the preparation of oyster mushroom, yoghurt formulation with three levels of supplementation, and evaluation of physicochemical, sensory, microbiological, and antimicrobial properties. Preliminary sensory acceptability involved 37 panelists, and LAB viability was assessed using total plate count on MRS agar. Inhibitory effect was evaluated against E. coli using the well diffusion assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test, and the E. coli inhibitory effect was analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. Oyster mushroom supplementation significantly increased total culturable LAB viability at YOM 3% and YOM 5%, whereas a significantly larger inhibition zone against E. coli was observed only at YOM 5%. These findings provide preliminary evidence that oyster mushroom supplementation may support the growth of the mixed starter culture and produce limited inhibitory activity against E. coli, although its strain-specific prebiotic effect and broader antimicrobial potential require further confirmation.

Physicochemical dynamics of purple sweet potato vinegar production with agarwood leaf infusion

Sin Yun Tan, Ying Ping Chang

APJMBB 34(3) (Special): 68-76

Article DOI: https://doi.org/10.35118/apjmbb.2026.034.3(Special).07

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This study selected a tuber substrate for vinegar production and evaluated the effect of agarwood leaf infusion during subsequent fermentation of the selected substrate. Purple sweet potato and cassava were first saccharified using Aspergillus oryzae; purple sweet potato yielded significantly more reducing sugar after saccharification (28.40%) and was therefore used for all subsequent alcoholic and acetic acid fermentations. Agarwood leaf infusion increased the total phenolic content during the 10-day acetification period without significantly changing final acetic acid concentration (4.536 mg/mL versus 4.302 mg/mL in the water-based control). The infusion also increased phytic acid concentration and altered the minor organic-acid profile, including detection of propionic acid at the start of acetification. Qualitative phytochemical screening showed comparable flavonoid, tannin and coumarin results between treatments; quinone was detected only in the final water-based sample. Therefore, purple sweet potato was suitable for this stepwise fermentation system, while agarwood leaf infusion modified total phenolic content and the organic-acid profile without impeding acetification. The reported final acetic acid concentrations remained below the Malaysian regulatory minimum for a commercially marketed vinegar, indicating that further process optimisation is required.