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Fulltext Establishment and analysis of the 3-dimensional (3D) spheroids generated from the Nasopharyngeal Carcinoma cell line HK1

Muniandy, Kalaivani, Sankar, Prabu Siva, Lian, Benedict Shi Xiang, Khoo, Alan Soo-Beng, Balakrishnan, Venugopal, Mohana-Kumaran, Nethia

Trop Life Sci Res, 2016;27(11):125-130.
MyJurnal

Spheroids have been shown to recapitulate the tumour in vivo with properties
such as the tumour microenvironment, concentration gradients, and tumour phenotype. As
such, it can serve as a platform for determining the growth and invasion behaviour pattern
of the cancer cells as well as be utilised for drug sensitivity assays; capable of exhibiting
results that are closer to what is observed in vivo compared to two-dimensional (2D) cell
culture assays. This study focused on establishing a three-dimensional (3D) cell culture
model using the Nasopharyngeal Carcinoma (NPC) cell line, HK1 and analysing its growth
and invasion phenotypes. The spheroids will also serve as a model to elucidate their
sensitivity to the chemotherapeutic drug, Flavopiridol. The liquid overlay method was
employed to generate the spheroids which was embedded in bovine collagen I matrix for
growth and invasion phenotypes observation. The HK1 cells formed compact spheroids
within 72 hours. Our observation from the 3 days experiments revealed that the spheroids
gradually grew and invaded into the collagen matrix, showing that the HK1 spheroids are
capable of growth and invasion. Progressing from these experiments, the HK1 spheroids
were employed to perform a drug sensitivity assay using the chemotherapeutic drug,
Flavopiridol. The drug had a dose-dependent inhibition on spheroid growth and invasion.
Isolation and genotyping of Toxoplasma gondii from free-range ducks in Malaysia

Puvanesuaran VR, Noordin R, Balakrishnan V

Avian Dis, 2013 Mar;57(1):128-32.
PMID: 23678741

Toxoplasma gondii is a parasitic protozoan that infects nearly one-third of humans. The present study was performed to isolate and genotype T. gondii from free-range ducks in Malaysia. Sera, heads, and hearts from 205 ducks were obtained from four states in Peninsular Malaysia, and 30 (14.63%) sera were found to be seropositive when assayed with the modified agglutination test (MAT > or = 1:6). All the positive samples were inoculated into mice, and T. gondii was successfully isolated from four individual duck samples (1.95%), which were initially found to be strongly seropositive (MAT > or = 1:24). The isolates were subjected to PCR-RFLP analysis, and two T. gondii strains were identified: type I and type II. This is the first reported study on the genetic characterization of T. gondii isolates from free-range farm animals in Southeast Asia.
Fulltext Genotyping of Toxoplasma gondii isolates from wild boars in Peninsular Malaysia

Puvanesuaran VR, Noordin R, Balakrishnan V

PLoS One, 2013;8(4):e61730.
PMID: 23613920 DOI: 10.1371/journal.pone.0061730

Toxoplasma gondii is a parasitic protozoan that infects nearly one-third of the world population. The present study was done to isolate and genotype T. gondii from wild boar from forests of Pahang, Malaysia. A total of 30 wild boars' blood, heads and hearts were obtained for this study and 30 (100.0%) were found to be seropositive when assayed with modified agglutination test (MAT ≥ 6). The positive samples were inoculated into mice and T. gondii was only isolated from samples that had strong seropositivity (MAT ≥ 1:24).The isolates were subjected to PCR-RFLP analysis and all the Peninsular Malaysia isolates of T. gondii are of clonal type I.
THICAPA modulates amyloid precursor protein processing and alleviates oxidative stress in fibroblasts derived from patients with Familial Alzheimer's disease

Thangeswaran D, Shamsuddin S, Balakrishnan V

Mol Biol Rep, 2025 Aug 19;52(1):835.
PMID: 40828210 DOI: 10.1007/s11033-025-10930-4

BACKGROUND: Familial Alzheimer's disease (fAD) is a hereditary disease that develops at an unusually early age. The deposition of toxic amyloid-beta (Aβ) is a hallmark of fAD. Despite their genetic origin and increasing prevalence, no effective drugs currently exist. THICAPA, a novel compound containing a tetrahydroisoquinoline group of amines, is naturally found in the brain and food and has diverse medicinal properties. However, its potential role in modulating amyloidogenesis in patients with fAD has not yet been explored.
METHODS AND RESULTS: We investigated the effects of THICAPA on the amyloid precursor protein (APP) processing pathway in fibroblasts derived from patients with fAD. The in vitro cytotoxicity assay revealed no significant THICAPA cytotoxicity in fAD (AG06840) or healthy fibroblast cell lines (GM05879). Aβ scavenging assay revealed that 50 µM THICAPA potentially scavenged aged Aβ42 oligomers in the healthy fibroblast line. Gene and protein expression analyses revealed reduced APP expression and mature/immature APP expression ratio, BACE1 and presenilin 1 downregulation, and ADAM10 upregulation. Protein quantification revealed a significant reduction in C-terminal fragment beta, soluble APP (sAPP)β, and Aβ42/40 ratio in the amyloidogenic pathway and elevated sAPPα in the non-amyloidogenic pathway. Moreover, reactive oxygen species detection indicated that THICAPA reduced ROS production in fibroblasts from patients with fAD by 41.63%, although its intrinsic antioxidant properties were modest.
CONCLUSIONS: THICAPA attenuates amyloidogenesis and upregulates the non-amyloidogenic pathway while alleviating ROS production. These findings suggest that THICAPA is a potential therapeutic candidate for treating fAD.
Fulltext Key Molecular Events in Cervical Cancer Development

Balasubramaniam SD, Balakrishnan V, Oon CE, Kaur G

Medicina (Kaunas), 2019 Jul 17;55(7).
PMID: 31319555 DOI: 10.3390/medicina55070384

Cervical cancer is the fourth most common cancer among women. Infection by high-risk human papillomavirus (HPV) is the main aetiology for the development of cervical cancer. Infection by high-risk human papillomavirus (HPV) and the integration of the HPV genome into the host chromosome of cervical epithelial cells are key early events in the neoplastic progression of cervical lesions. The viral oncoproteins, mainly E6 and E7, are responsible for the initial changes in epithelial cells. The viral proteins inactivate two main tumour suppressor proteins, p53, and retinoblastoma (pRb). Inactivation of these host proteins disrupts both the DNA repair mechanisms and apoptosis, leading to rapid cell proliferation. Multiple genes involved in DNA repair, cell proliferation, growth factor activity, angiogenesis, as well as mitogenesis genes become highly expressed in cervical intraepithelial neoplasia (CIN) and cancer. This genomic instability encourages HPV-infected cells to progress towards invasive carcinoma. The key molecular events involved in cervical carcinogenesis will be discussed in this review.
The potential applications of T cell receptor (TCR)-like antibody in cervical cancer immunotherapy

Dass SA, Selva Rajan R, Tye GJ, Balakrishnan V

Hum Vaccin Immunother, 2021 09 02;17(9):2981-2994.
PMID: 33989511 DOI: 10.1080/21645515.2021.1913960

Cervical cancer is ranked as the fourth most common cancer in women worldwide. Monoclonal antibody has created a new dimension in the immunotherapy of many diseases, including cervical cancer. The antibody's ability to target various aspects of cervical cancer (oncoviruses, oncoproteins, and signaling pathways) delivers a promising future for efficient immunotherapy. Besides, technologies such as hybridoma and phage display provide a fundamental platform for monoclonal antibody generation and create the opportunity to generate novel antibody classes including, T cell receptor (TCR)-like antibody. In this review, the current immunotherapy strategies for cervical cancer are presented. We have also proposed a novel concept of T cell receptor (TCR)-like antibody and its potential applications for enhancing cervical cancer therapeutics. Finally, the possible challenges in TCR-like antibody application for cervical cancer therapeutics have been addressed, and strategies to overcome the challenges have been highlighted to maximize the therapeutic benefits.
Fulltext Minichromosome Maintenance Complex (MCM) Genes Profiling and MCM2 Protein Expression in Cervical Cancer Development

Kaur G, Balasubramaniam SD, Lee YJ, Balakrishnan V, Oon CE

Asian Pac J Cancer Prev, 2019 Oct 01;20(10):3043-3049.
PMID: 31653153 DOI: 10.31557/APJCP.2019.20.10.3043

OBJECTIVE: Minichromosome maintenance complex (MCM) proteins are essential for the process of DNA replication and cell division. This study aimed to evaluate MCM genes expression profiles and MCM2 protein in HPV-associated cervical carcinogenesis.
METHODOLOGY: MCM2, 4, 5 and 7 genes expression profiles were evaluated in three cervical tissue samples each of normal cervix, human papillomavirus (HPV)-infected low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL) and squamous cell carcinoma (SCC), using Human Transcriptome Array 2.0 and validated by nCounter® PanCancer Pathway NanoString Array. Immunohistochemical expression of MCM2 protein was semi-quantitatively assessed by histoscore in tissue microarrays containing 9 cases of normal cervix, 10 LSIL, 10 HSIL and 42 cases of SCC.
RESULTS: MCM2, 4, 5 and 7 genes expressions were upregulated with increasing fold change during the progression from LSIL to HSIL and the highest in SCC. MCM2 gene had the highest fold change in SCC compared to normal cervix. Immunohistochemically, MCM2 protein was localised in the nuclei of basal cells of normal cervical epithelium and dysplastic-neoplastic cells of CIN and SCC. There was a significant difference in MCM2 protein expression between the histological groups (P = 0.039), and histoscore was the highest in HSIL compared to normal cervix (P = 0.010).
CONCLUSION: The upregulation of MCM genes expressions in cervical carcinogenesis reaffirms MCM as a proliferative marker in DNA replication pathway, whereby proliferation of dysplastic and cancer cells become increasingly dysregulated and uncontrolled. A strong expression of MCM2 protein in HSIL may aid as a concatenated screening tool in detecting pre-cancerous cervical lesions.
Comparative transcriptomic profiling in HPV-associated cervical carcinogenesis: Implication of MHC class II and immunoglobulin heavy chain genes

Balasubramaniam SD, Wong KK, Oon CE, Balakrishnan V, Kaur G

Life Sci, 2020 Sep 01;256:118026.
PMID: 32615187 DOI: 10.1016/j.lfs.2020.118026

AIM: We aimed to determine the biological processes and pathways involved in cervical carcinogenesis associated with high-risk human papillomavirus (HPV) infection.
MATERIALS AND METHODS: Total RNA was extracted from three formalin-fixed paraffin-embedded (FFPE) samples each of normal cervix, HPV-infected low-grade squamous intraepithelial lesion (LSIL), high-grade SIL (HSIL) and squamous cell carcinoma (SCC). Transcriptomic profiling by microarrays was conducted followed by downstream Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses.
RESULTS: We examined the difference in GOs enriched for each transition stage from normal cervix to LSIL, HSIL, and SCC, and found 307 genes to be differentially expressed. In the transition from normal cervix to LSIL, the extracellular matrix (ECM) genes were significantly downregulated. The MHC class II genes were significantly upregulated in the LSIL to HSIL transition. In the final transition from HSIL to SCC, the immunoglobulin heavy locus genes were significantly upregulated and the ECM pathway was implicated.
CONCLUSION: Deregulation of the immune-related genes including MHC II and immunoglobulin heavy chain genes were involved in the transitions from LSIL to HSIL and SCC, suggesting immune escape from host anti-tumour response. The extracellular matrix plays an important role during the early and late stages of cervical carcinogenesis.
Fulltext Isolation and Characterization of Magnetic Oil Palm Empty Fruits Bunch Cellulose Nanofiber Composite as a Bio-Sorbent for Cu(II) and Cr(VI) Removal

Khalid AM, Hossain MS, Ismail N, Khalil NA, Balakrishnan V, Zulkifli M, et al.

Polymers (Basel), 2020 Dec 30;13(1).
PMID: 33396583 DOI: 10.3390/polym13010112

In the present study, magnetic oil palm empty fruits bunch cellulose nanofiber (M-OPEFB-CNF) composite was isolated by sol-gel method using cellulose nanofiber (CNF) obtained from oil palm empty fruits bunch (OPEFB) and Fe3O4 as magnetite. Several analytical methods were utilized to characterize the mechanical, chemical, thermal, and morphological properties of the isolated CNF and M-OPEFB-CNF. Subsequently, the isolated M-OPEFB-CNF composite was utilized for the adsorption of Cr(VI) and Cu(II) from aqueous solution with varying parameters, such as pH, adsorbent doses, treatment time, and temperature. Results showed that the M-OPEFB-CNF as an effective bio-sorbent for the removal of Cu(II) and Cr(VI) from aqueous solution. The adsorption isotherm modeling revealed that the Freundlich equation better describes the adsorption of Cu(II) and Cr(VI) on M-OPEFB-CNF composite. The kinetics studies revealed the pseudo-second-order kinetics model was a better-described kinetics model for the removal of Cu(II) and Cr(VI) using M-OPEFB-CNF composite as bio-sorbent. The findings of the present study showed that the M-OPEFB-CNF composite has the potential to be utilized as a bio-sorbent for heavy metals removal.
Fulltext Effect of Supporting Background Electrolytes on the Nanostructure Morphologies and Electrochemical Behaviors of Electrodeposited Gold Nanoparticles on Glassy Carbon Electrode Surfaces

Zakaria ND, Omar MH, Ahmad Kamal NN, Abdul Razak K, Sönmez T, Balakrishnan V, et al.

ACS Omega, 2021 Sep 28;6(38):24419-24431.
PMID: 34604624 DOI: 10.1021/acsomega.1c02670

Electrodeposition is an electrochemical method employed to deposit stable and robust gold nanoparticles (AuNPs) on electrode surfaces for creating chemically modified electrodes (CMEs). The use of several electrodeposition techniques with different experimental parameters allow in obtaining various surface morphologies of AuNPs deposited on the electrode surface. By considering the electrodeposition of AuNPs in various background electrolytes could play an important strategy in finding the most suitable formation of the electrodeposited AuNP films on the electrode surface. This is because different electrode roughnesses can have different effects on the electrochemical activities of the modified electrodes. Thus, in this study, the electrodeposition of AuNPs onto the glassy carbon (GC) electrode surfaces in various aqueous neutral and acidic electrolytes was achieved by using the cyclic voltammetry (CV) technique with no adjustable CV parameters. Then, surface morphologies and electrochemical activities of the electrodeposited AuNPs were investigated using scanning electron microscopy (SEM), atomic force microscopy (AFM), CV, and electrochemical impedance spectroscopy (EIS). The obtained SEM and 3D-AFM images show that AuNPs deposited at the GC electrode prepared in NaNO3 solution form a significantly better, uniform, and homogeneous electrodeposited AuNP film on the GC electrode surface with nanoparticle sizes ranging from ∼36 to 60 nm. Meanwhile, from the electrochemical performances of the AuNP-modified GC electrodes, characterized by using a mixture of ferricyanide and ferrocyanide ions [Fe(CN6)3-/4-], there is no significant difference observed in the case of charge-transfer resistances (R ct) and heterogeneous electron-transfer rate constants (k o), although there are differences in the surface morphologies of the electrodeposited AuNP films. Remarkably, the R ct values of the AuNP-modified GC electrodes are lower than those of the bare GC electrode by 18-fold, as the R ct values were found to be ∼6 Ω (p < 0.001, n = 3). This has resulted in obtaining k o values of AuNP-modified GC electrodes between the magnitude of 10-2 and 10-3 cm s-1, giving a faster electron-transfer rate than that of the bare GC electrode (10-4 cm s-1). This study confirms that using an appropriate supporting background electrolyte plays a critical role in preparing electrodeposited AuNP films. This approach could lead to nanostructures with a more densely, uniformly, and homogeneously electrodeposited AuNP film on the electrode surfaces, albeit utilizing an easy and simple preparation method.
Fulltext Treatment of clinical solid waste using a steam autoclave as a possible alternative technology to incineration

Hossain MS, Balakrishnan V, Rahman NN, Sarker MZ, Kadir MO

Int J Environ Res Public Health, 2012 Mar;9(3):855-67.
PMID: 22690168 DOI: 10.3390/ijerph9030855

A steam autoclave was used to sterilize bacteria in clinical solid waste in order to determine an alternative to incineration technology in clinical solid waste management. The influence of contact time (0, 5, 15, 30 and 60 min) and temperature (111 °C, 121 °C and 131 °C) at automated saturated steam pressure was investigated. Results showed that with increasing contact time and temperature, the number of surviving bacteria decreased. The optimum experimental conditions as measured by degree of inactivation of bacteria were 121 °C for 15 minutes (min) for Gram negative bacteria, 121 °C and 131 °C for 60 and 30 min for Gram positive bacteria, respectively. The re-growth of bacteria in sterilized waste was also evaluated in the present study. It was found that bacterial re-growth started two days after the inactivation. The present study recommends that the steam autoclave cannot be considered as an alternative technology to incineration in clinical solid waste management.
Fulltext Infectious risk assessment of unsafe handling practices and management of clinical solid waste

Hossain MS, Rahman NN, Balakrishnan V, Puvanesuaran VR, Sarker MZ, Kadir MO

Int J Environ Res Public Health, 2013 Jan 31;10(2):556-67.
PMID: 23435587 DOI: 10.3390/ijerph10020556

The present study was undertaken to determine the bacterial agents present in various clinical solid wastes, general waste and clinical sharp waste. The waste was collected from different wards/units in a healthcare facility in Penang Island, Malaysia. The presence of bacterial agents in clinical and general waste was determined using the conventional bacteria identification methods. Several pathogenic bacteria including opportunistic bacterial agent such as Pseudomonas aeruginosa, Salmonella spp., Klebsiella pneumoniae, Serratia marcescens, Acinetobacter baumannii, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus pyogenes were detected in clinical solid wastes. The presence of specific pathogenic bacterial strains in clinical sharp waste was determined using 16s rDNA analysis. In this study, several nosocomial pathogenic bacteria strains of Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Lysinibacillus sphaericus, Serratia marcescens, and Staphylococcus aureus were detected in clinical sharp waste. The present study suggests that waste generated from healthcare facilities should be sterilized at the point of generation in order to eliminate nosocomial infections from the general waste or either of the clinical wastes.
Fulltext The COVID-19/Tuberculosis Syndemic and Potential Antibody Therapy for TB Based on the Lessons Learnt From the Pandemic

Dass SA, Balakrishnan V, Arifin N, Lim CSY, Nordin F, Tye GJ

Front Immunol, 2022;13:833715.
PMID: 35242137 DOI: 10.3389/fimmu.2022.833715

2020 will be marked in history for the dreadful implications of the COVID-19 pandemic that shook the world globally. The pandemic has reshaped the normality of life and affected mankind in the aspects of mental and physical health, financial, economy, growth, and development. The focus shift to COVID-19 has indirectly impacted an existing air-borne disease, Tuberculosis. In addition to the decrease in TB diagnosis, the emergence of the TB/COVID-19 syndemic and its serious implications (possible reactivation of latent TB post-COVID-19, aggravation of an existing active TB condition, or escalation of the severity of a COVID-19 during TB-COVID-19 coinfection), serve as primary reasons to equally prioritize TB. On a different note, the valuable lessons learnt for the COVID-19 pandemic provide useful knowledge for enhancing TB diagnostics and therapeutics. In this review, the crucial need to focus on TB amid the COVID-19 pandemic has been discussed. Besides, a general comparison between COVID-19 and TB in the aspects of pathogenesis, diagnostics, symptoms, and treatment options with importance given to antibody therapy were presented. Lastly, the lessons learnt from the COVID-19 pandemic and how it is applicable to enhance the antibody-based immunotherapy for TB have been presented.
Optimizing supercritical carbon dioxide in the inactivation of bacteria in clinical solid waste by using response surface methodology

Hossain MS, Nik Ab Rahman NN, Balakrishnan V, Alkarkhi AF, Ahmad Rajion Z, Ab Kadir MO

Waste Manag, 2015 Apr;38:462-73.
PMID: 25636860 DOI: 10.1016/j.wasman.2015.01.003

Clinical solid waste (CSW) poses a challenge to health care facilities because of the presence of pathogenic microorganisms, leading to concerns in the effective sterilization of the CSW for safe handling and elimination of infectious disease transmission. In the present study, supercritical carbon dioxide (SC-CO2) was applied to inactivate gram-positive Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis, and gram-negative Escherichia coli in CSW. The effects of SC-CO2 sterilization parameters such as pressure, temperature, and time were investigated and optimized by response surface methodology (RSM). Results showed that the data were adequately fitted into the second-order polynomial model. The linear quadratic terms and interaction between pressure and temperature had significant effects on the inactivation of S. aureus, E. coli, E. faecalis, and B. subtilis in CSW. Optimum conditions for the complete inactivation of bacteria within the experimental range of the studied variables were 20 MPa, 60 °C, and 60 min. The SC-CO2-treated bacterial cells, observed under a scanning electron microscope, showed morphological changes, including cell breakage and dislodged cell walls, which could have caused the inactivation. This espouses the inference that SC-CO2 exerts strong inactivating effects on the bacteria present in CSW, and has the potential to be used in CSW management for the safe handling and recycling-reuse of CSW materials.
Fulltext Effectiveness of Booster Vaccinations on the Control of COVID-19 during the Spread of Omicron Variant in Malaysia

Wong MTJ, Dhaliwal SS, Balakrishnan V, Nordin F, Norazmi MN, Tye GJ

Int J Environ Res Public Health, 2023 Jan 16;20(2).
PMID: 36674401 DOI: 10.3390/ijerph20021647

(1) Background: The assessment of vaccine effectiveness against the Omicron variant is vital in the fight against COVID-19, but research on booster vaccine efficacy using nationwide data was lacking at the time of writing. This study investigates the effectiveness of booster doses on the Omicron wave in Malaysia against COVID-19 infections and deaths; (2) Methods: This study uses nationally representative data on COVID-19 from 1 January to 31 March 2022, when the Omicron variant was predominant in Malaysia. Daily new infections, deaths, ICU utilization and Rt values were compared. A screening method was used to predict the vaccine effectiveness against COVID-19 infections, whereas logistic regression was used to estimate vaccine effectiveness against COVID-19-related deaths, with efficacy comparison between AZD1222, BNT162b2 and CoronaVac; (3) Results: Malaysia's Omicron wave started at the end of January 2022, peaking on 5 March 2022. At the time of writing, statistics for daily new deaths, ICU utilization, and effective reproductive values (Rt) were showing a downtrend. Boosted vaccination is 95.4% (95% CI: 95.4, 95.4) effective in curbing COVID-19 infection, compared to non-boosted vaccination, which is 87.2% (95% CI: 87.2, 87.2). For symptomatic infection, boosted vaccination is 97.4% (95% CI: 97.4, 97.4) effective, and a non-boosted vaccination is 90.9% (95% CI: 90.9, 90.9). Against COVID-19-related death, boosted vaccination yields a vaccine effectiveness (VE) of 91.7 (95% CI: 90.6, 92.7) and full vaccination yields a VE of 65.7% (95% CI: 61.9, 69.1). Looking into the different vaccines as boosters, AZD1222 is 95.2% (CI 95%: 92.7, 96.8) effective, BNT162b2 is 91.8% (CI 95%: 90.7, 92.8) effective and CoronaVac is 88.8% (CI 95%: 84.9, 91.7) effective against COVID-19 deaths. (4) Conclusions: Boosters are effective in increasing protection against COVID-19, including the Omicron variant. Given that the VE observed was lower, CoronaVac recipients are encouraged to take boosters due to its lower VE.
Fulltext Chimeric antigen receptor-natural killer cell therapy: current advancements and strategies to overcome challenges

Kong JC, Sa'ad MA, Vijayan HM, Ravichandran M, Balakrishnan V, Tham SK, et al.

Front Immunol, 2024;15:1384039.
PMID: 38726000 DOI: 10.3389/fimmu.2024.1384039

Chimeric antigen receptor-natural killer (CAR-NK) cell therapy is a novel immunotherapy targeting cancer cells via the generation of chimeric antigen receptors on NK cells which recognize specific cancer antigens. CAR-NK cell therapy is gaining attention nowadays owing to the ability of CAR-NK cells to release potent cytotoxicity against cancer cells without side effects such as cytokine release syndrome (CRS), neurotoxicity and graft-versus-host disease (GvHD). CAR-NK cells do not require antigen priming, thus enabling them to be used as "off-the-shelf" therapy. Nonetheless, CAR-NK cell therapy still possesses several challenges in eliminating cancer cells which reside in hypoxic and immunosuppressive tumor microenvironment. Therefore, this review is envisioned to explore the current advancements and limitations of CAR-NK cell therapy as well as discuss strategies to overcome the challenges faced by CAR-NK cell therapy. This review also aims to dissect the current status of clinical trials on CAR-NK cells and future recommendations for improving the effectiveness and safety of CAR-NK cell therapy.
Advancement in diagnostic approaches for latent tuberculosis: distinguishing recent from remote infections

Ding YE, Wong MTJ, Norazmi MN, Balakrishnan V, Tye GJ

One Health Outlook, 2025 Apr 10;7(1):19.
PMID: 40205610 DOI: 10.1186/s42522-025-00144-w

Tuberculosis (TB) remains as a significant global health threat to date, with latent TB infection (LTBI) serving as a major reservoir for future active disease cases. A practical approach to an effective control and eradication of TB hence, requires an explicit identification of infected patient whom are at high risk of progressing from latent to active TB, particularly in those recently infected individuals. Current diagnostic tools however, including Tuberculin Skin Test and Interferon-Gamma Release Assays, are still lacking for their ability to critically distinguish between recent and remote infections, leading to insufficiency in optimizing targeted preventive treatment strategies. This review examines the limitations of current diagnostic tools and explores novel biomarkers to enhance distinction within the infection timeline in LTBI diagnostics. Advancement in immune profiling, dormancy antigen, along with molecular and transcriptomic approaches holds great promise to develop a diagnostic tools with better accuracy to differentiate recent from remote infections, thereby optimizing targeted interventions to improve TB control strategies. These underscores the need for further research into these emerging diagnostic tools to facilitate an effective public health strategies and contribute to the united efforts in End TB Strategy.
Fulltext Recycling Waste Cotton Cloths for the Isolation of Cellulose Nanocrystals: A Sustainable Approach

Mohamed SH, Hossain MS, Mohamad Kassim MH, Ahmad MI, Omar FM, Balakrishnan V, et al.

Polymers (Basel), 2021 Feb 19;13(4).
PMID: 33669623 DOI: 10.3390/polym13040626

There is an interest in the sustainable utilization of waste cotton cloths because of their enormous volume of generation and high cellulose content. Waste cotton cloths generated are disposed of in a landfill, which causes environmental pollution and leads to the waste of useful resources. In the present study, cellulose nanocrystals (CNCs) were isolated from waste cotton cloths collected from a landfill. The waste cotton cloths collected from the landfill were sterilized and cleaned using supercritical CO2 (scCO2) technology. The cellulose was extracted from scCO2-treated waste cotton cloths using alkaline pulping and bleaching processes. Subsequently, the CNCs were isolated using the H2SO4 hydrolysis of cellulose. The isolated CNCs were analyzed to determine the morphological, chemical, thermal, and physical properties with various analytical methods, including attenuated total reflection-Fourier transform-infrared spectroscopy (ATR-FTIR), field-emission scanning electron microscopy (FE-SEM), energy-filtered transmission electron microscopy (EF-TEM), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). The results showed that the isolated CNCs had a needle-like structure with a length and diameter of 10-30 and 2-6 nm, respectively, and an aspect ratio of 5-15, respectively. Additionally, the isolated CNCs had a high crystallinity index with a good thermal stability. The findings of the present study revealed the potential of recycling waste cotton cloths to produce a value-added product.
Fulltext Influence of Fresh Palm Fruit Sterilization in the Production of Carotenoid-Rich Virgin Palm Oil

Hassan NSM, Hossain MS, Balakrishnan V, Zuknik MH, Mustaner M, Easa AM, et al.

Foods, 2021 Nov 17;10(11).
PMID: 34829117 DOI: 10.3390/foods10112838

Palm oil is known to be rich in carotenoids and other phytonutrients. However, the carotenoids and phytonutrients degrade due to high heat sterilization of oil palm fruits. The present study was conducted to produce carotenoid-rich virgin palm oil (VPO) using cold-press extraction. Herein, the influence of sterilization of oil palm fresh fruits in the production of cold-pressed VPO was determined with varying sterilization temperatures, times, and amounts of palm fruits in sterilization. The experimental sterilization conditions were optimized using response surface methodology (RSM) based on the maximum VPO yield and minimum FFAs in cold-pressed VPO. The optimal sterilization experimental conditions of oil palm fruits were determined to be a sterilization temperature of 62 °C, a time of 90 min, and an amount of oil palm fruits of 8 kg. Under these experimental conditions, the maximum cold-pressed VPO yield and the minimal content of free fatty acids (FFAs) obtained were 27.94 wt.% and 1.32 wt.%, respectively. Several analytic methods were employed to determine cold-pressed VPO quality and fatty acids compositions and compared with the crude palm oil. It was found that cold-pressed VPO contains higher carotenoids (708 mg/g) and unsaturated fatty acids compared with the carotenoid (343 mg/g) and fatty acid compositions in CPO. The findings of the present study reveal that the sterilization temperature potentially influences the carotenoid and nutrient contents in VPO; therefore, the optimization of the sterilization conditions is crucial to producing carotenoid- and phytonutrient-rich VPO.
Electrochemical and imaging evaluations of electrochemically activated screen-printed gold electrodes

Zakaria ND, Salih IL, Hamzah HH, Sönmez T, Omar MH, Nor NM, et al.

Analyst, 2024 Nov 04;149(22):5401-5410.
PMID: 39373774 DOI: 10.1039/d4an00990h

Sulfuric acid is commonly used to electrochemically activate gold electrodes in a variety of electrochemical applications. This work provides the first evaluations of the electrochemical behaviors and a 3D image of an activated screen-printed gold electrode (SPGE, purchased commercially) through electrochemical and imaging analyses. The activated SPGE surface appears rougher than the unactivated SPGE surface when viewed through microtopography images using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Nevertheless, the roughened microscopy structure does not exhibit any substantial changes in roughness factor for the activated SPGE, as indicated by capacitive current analyses. The significant improvement in electrochemical responsiveness of the activated SPGE is mainly attributed to the presence of surface pores created in the microscopic structure as a result of gold oxide layer formation. The presence of surface pores on the activated surface has significantly improved its conductivity by 10-fold. As a result, electron transfer kinetics and mass transports of the activated SPGE are greatly improved. The results presented in this work indicate that the surface of the activated SPGE greatly increased its intrinsic surface pores, and conductivity of the electrode surface and uncovered the electrocatalytic active sites. This significantly improves the activated SPGE's performance in electrochemical applications such as oxygen reduction reaction (ORR). An activated SPGE considerably enhanced limiting current density as well as ∼172 mV versus Ag shifted onset potential to more positive potentials compared to unactivated SPGE.

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