Background: Bacterial biofilms within the root canal system are a major contributor in the failure of endodontic treatment, primarily attributed to the presence of significant pathogens such as Enterococcus faecalis (E. faecalis), Fusobacterium nucleatum (F. nucleatum), and Porphyromonas gingivalis (P. gingivalis), which demonstrate considerable virulence, antimicrobial resistance, and the capacity to penetrate dentinal tubules. Conventional irrigants such sodium hypochlorite (NaOCl) and chlorhexidine gluconate (CHX) have limitations including cytotoxicity, inadequate biofilm eradication, and adverse responses including tissue irritation and precipitate formation. Quaternary ammonium silane (codename- K21), a functionalised organosilicon compound with antibacterial and anti-inflammatory properties, not only disrupts microbial biofilms but also preserves compatibility with host tissue components, including macrophages. Comprehensive research is warranted to confirm K21's therapeutic benefits since it has potential to block Sortase Av(SrtA) and matrix metalloproteinases (MMPs), enzymes engaged in bacterial adherence and tissue breakdown. Objectives: The aim of this research was to assess K21's antibacterial and anti-inflammatory performance as an endodontic irrigant. Among specific objectives were comparing 1.00% K21 with 0.5% K21, 2.00% CHX, 6.00% NaOCl, and saline on single-species (E. faecalis, F. nucleatum and P. gingivalis respectively) and dual-species (F. nucleatum and P. gingivalis) biofilms at 200 μm and 400 μm dentinal tubule depths. Additionally evaluated in the study were K21's effects on hPDLSCs survival as well as RAW 264.7 macrophage viability and anti-inflammatory (M2) polarization. Moreover, molecular simulations were performed to assess the interaction of K21 with SrtA and MMP-9 molecular structures. While the null hypothesis predicted no significant differences, the hypotheses postulated that 1.00% K21 would show better biofilm reduction, higher macrophage and hPDLSCs viability, and greater anti-inflammatory benefits than conventional irrigants. Methods: This multimodal investigation utilized a combination of in vitro assays and imaging techniques. Biofilms of E. faecalis, F. nucleatum, and P. gingivalis were cultured on dentin specimens and exposed to different irrigants: saline, 6.00% NaOCl, 2.00% CHX, 0.5%K21, and 1.00% K21. Colony-forming unit (CFU) counts were used to assess bacterial viability, while structural changes were analyzed via scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Raman spectroscopy was employed to identify chemical alterations in post-treatment biofilm components. Cell viability was evaluated using the MTT assay and mitochondrial staining in RAW264.7 macrophages, while the alamar blue assay was utilised for hPDLSCs. Additionally, the expression of cytokines (TNF-α, IL-1β, TGF-β, VEGF-A) associated with macrophage polarization was evaluated using Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) and gene expression analysis to determine the immunomodulatory impact of all the irrigants. Molecular simulations were carried out inside the Schrödinger Drug Discovery Suite using the Desmond package. Molecular simulations analyzed K21’s binding to MMP-9 and SrtA molecular structures with stability assessed via root mean square deviation (RMSD) and binding free energy (MM-GBSA). Statistical analysis used ANOVA with Tukey’s post hoc test (p < 0.05) and Pearson’s correlation for depth-dependent efficacy. Results: The CFU analysis revealed that K21, especially at a 1.00% concentration, consistently showed enhanced antibacterial efficacy relative to 6.00% NaOCl and was frequently comparable to or more effective than 2.00% CHX against E. faecalis, F. nucleatum, P. gingivalis, and dual-species biofilms. For CFU findings of E. faecalis, at a depth of 200 μm and a duration of 1-minute, 1.00% K21 exhibited the lowest CFU at 1.358 ± 0.193, in contrast to 0.5% K21 (3.453 ± 0.205), 2.00% CHX (4.65 ± 0.093), 6.00% NaOCl (5.64 ± 0.108), and saline (9.63 ± 0.031), with statistically significant differences (p < 0.001). At 400 μm and 1-minute, similar trends were observed. In 5-minute exposures, 1.00% K21 consistently exhibited the lowest CFUs (0.9687–0.9924), significantly surpassing 2.00% CHX and 6.00% NaOCl at both 200 μm and 400 μm (p < 0.001). For F. nucleatum biofilm, 1.00% K21 exhibited the lowest CFUs (2.1199 ± 0.20274 at 200 μm/5 min and 2.1782 ± 0.55736 at 400 μm/5 min), demonstrating significant reductions compared to 6.00% NaOCl (p < 0.001) and comparable efficacy to 2.00% CHX. At 1-minute application, CFU counts were markedly diminished by K21 group compared to 6.00% NaOCl. For P. gingivalis, at 200 μm/1 min, 1.00% K21 (3.9831 ± 0.10972) and 0.5%K21 were significantly better than 6.00% NaOCl (5.1798 ± 0.39327) and were comparable to 2.00% CHX (4.0792 ± 0.10074). At 400 μm/1 min and 5 min, all test irrigants demonstrated effectiveness relative to saline; nevertheless, 1.00% K21 consistently exhibited the lowest CFU counts with no statistically significant difference from 2.00% CHX. In dual-species biofilm, 1.00% K21 demonstrated significant reductions compared to saline at both 200 μm and 400 μm for 1 and 5-minute treatments. At 200 μm/5 min, 1.00% K21 (4.6173 ± 0.37708) and 0.5%K21 (4.7104 ± 0.34656) demonstrated superior efficacy compared to 6.00% NaOCl (5.1432 ± 0.48289). 1.00% K21 was comparable to 2.00% CHX (p > 0.05) and significantly better than saline (p < 0.001) and 6.00% NaOCl (p = 0.046). Morphological analyses utilising SEM and TEM corroborated the CFU findings, demonstrating that 1.00% K21 induced significant membrane rupture, cytoplasmic leakage, and biofilm matrix disintegration across all the three species. The 1.00% K21 treated groups exhibited the most pronounced intensity deterioration, and Raman spectroscopy examination indicated notable spectrum shifts in the 480–490 cm⁻¹ range, so corroborating the hypothesis of heightened structural disruption of bacterial components. For macrophage viability, all groups showed cell viability above 80%. While 0.5% and 1.00% K21 showed higher viability (87.17% and 87.4%), viability reduced for 6.00% NaOCl (82.42%) and 2.00% CHX (86.08%). K21 groups were noticeably better than 6.00% NaOCl, although all test groups had lowered viability relative to saline (p<0.05). Fluorescence imaging depicted strong, homogeneous mitochondrial signals maintained by saline and 0.5% K21 that indicated healthy, viable cells. The 1.00% K21 group maintained general function while only modestly altered mitochondrial structure. While 6.00% NaOCl produced severe cytotoxicity, low cell density, and weak fluorescence, 2.00% CHX exhibited variable effects with some mitochondrial fragmentation. At a concentration of 0.5%, K21 demonstrated superior mitochondrial preservation compared to 2.00% CHX and 6.00% NaOCl. SEM and TEM analysis indicated that macrophages treated with saline exhibited normal integrity and morphology. K21 exhibited electron-dense inclusions at elevated concentrations, signifying stress, but preserving cell morphology with slight surface alterations. The 6.00% NaOCl treatment caused severe structural damage, membrane fragmentation, and cytoplasmic debris; 2.00% CHX treated cells showed surface shrinkage and internal aggregates. K21 group displayed generally better cellular preservation than CHX and NaOCl groups. Gene expression analysis demonstrated that K21 induced modest upregulation of pro-inflammatory cytokines (TNF-α: ~4.24–4.26; IL-1β: ~1.43–1.89), whereas 2.00% CHX and 6.00% NaOCl strongly upregulated both TNF-α (~19.38 and 19.30) and IL-1β (~2.30 and 1.89). Conversely, K21-treated groups exhibited significantly higher anti-inflammatory markers, with VEGF-A reaching 58.93 ± 0.16 for 1.00% K21 and TGF-β at 6.51 ± 0.11, substantially exceeding levels observed with CHX (TGF-β: 6.31 ± 0.66) and NaOCl. Overall, K21 demonstrated a balanced immunomodulatory profile characterized by reduced inflammatory activation and enhanced pro-healing capacity compared to conventional irrigants. At 30 minutes of exposure, the viability of human periodontal ligament stem cells (hPDLSCs) remained approximately close to 80% for all tested irrigating agents. Specifically, 1.00% K21 demonstrated a viability of 81.13%, 0.5% K21 at 80.13%, CHX at 80.16%, and NaOCl at 79.34%. Saline showed the highest cell viability at 98.48%. High viability was indicated by SEM and TEM studies of hPDLSCs showing intact morphology and membrane integrity from saline-treated cells. Severe structural damage, membrane lysis, and cytoplasmic disintegration were produced by 6.00% NaOCl. Moderate membrane distortion and vacuolation caused by 2.00% CHX indicated cytotoxic effects. Although 1.00% K21 showed some structural changes while 0.5%K21 maintained better integrity. TEM verified that although K21 caused subcellular changes, it maintained membrane continuity, hence indicating better biocompatibility than 2.00% CHX and 6.00% NaOCl. The K21 group shows a specific pocket binding on several MMPs and specific SrtA structures, according to molecular simulation, which generates a classic clouting effect. This can prevent SrtA enzymes and MMPs from acting catalytically, avoiding structural changes in dentin. Conclusions The 1.00% K21 group demonstrated superior antibacterial efficacy against E. faecalis, F. nucleatum, and P. gingivalis in dentinal tubules at 200 μm and 400 μm depths compared to 6.00% NaOCl and saline, with performance comparable to 0.5% K21 and 2.00% CHX. In dual-species biofilms, 1.00% K21 outperformed NaOCl and saline while showing equivalent results to CHX and 0.5% K21. It exhibited significantly enhanced biocompatibility and anti-inflammatory properties in RAW 264.7 macrophages and hPDLSCs compared to NaOCl, with results comparable to 0.5% K21 and CHX. K21 promoted M2 macrophage polarization and a balanced immune response. Molecular docking revealed specific binding affinity to MMPs and Sortase A, indicating a unique "clouting effect" that enhances antibacterial and regenerative capabilities. Overall, 1.00% K21 represents a promising biocompatible, multifunctional endodontic irrigant with superior antibacterial and anti-inflammatory potential. Keywords: antibacterial, human periodontal ligament stem cells, K21, macrophages, molecular simulation, quaternary ammonium silane.

Heavy metals and plastics pollutions in the environment are becoming more common due to their extensive usage in our everyday lives and improper waste management. Interaction between heavy metals with plastics has been shown to have negative effects to the ecosystem. The aim of this study was to investigate the influence of polyethylene (PE) microplastic (MP) and nanoplastic (NP) on the combined toxicity of copper (Cu) and cadmium (Cd) in microalgae. The effects of PEMP and NP on the toxicity of Cu and Cd were assessed based on the algal physiological responses such as photosynthetic pigment contents and oxidative stress response. Our results showed that after 10 days of exposure, Cu and Cd caused more growth inhibition to Raphidocelis subcapitata (CuIC50=32.32 mg/L, Cd IC50=23.25 mg/L) compared to Chlorella sp. TJ6-5 (CuIC50=34.77 mg/L, Cd IC50=36.45 mg/L). Combinations of Cu and Cd at various IC concentrations (IC10, IC25, IC50, IC90) displayed synergistic effects and caused higher observed toxicity to both algal species compared to the expected level of toxicity. Although PE MP and NP did not cause significant growth inhibition to both algae species at concentrations up to 2 g/L. However, presence of PE MP and NP did enhance the growth inhibition effect of Cu and Cd (at IC50) on both algal strains with higher lipid peroxidation and reactive oxygen species (ROS) production. In addition, our results showed the combined exposure to mixture of PE MP and NP and Cu and Cd can compromise the cellular antioxidant defence capacity, potentially leading to increased susceptibility to oxidative damage and cellular death. In conclusion, the presence of PE MP/NP enhance the toxicity of Cu and Cd to freshwater microalgae particularly in R. subcapitata and Chlorella sp., with species-specific responses and varying degrees of synergistic effects.

Background: Learner agency is recognised as a critical attribute in health professions education, influencing students’ motivation, engagement, and capacity for self-regulated learning. However, the relationship between assessment practices and learner agency in medical education remains underexplored. Objective: This study explored the perceptions and experiences of undergraduate medical students regarding assessment practices and their influence on the development of learner agency within the MBBS programme at IMU University. Methods: A qualitative phenomenological design using focus group discussions (FGDs) was employed to examine the experiences of 27 medical students across Years 1 to 4. A purposive sample of 21 females and 6 males from diverse backgrounds participated in seven online FGDs conducted via Microsoft Teams©. Each session included 3–5 students. Data was analysed using iterative transcript review, inductive coding, and thematic analysis. Results: Three main dimensions of learner agency—environmental, behavioural, and intrapersonal—emerged as central to students’ experiences. Continuous assessments and timely, constructive feedback supported engagement and self-regulation. However, variability in feedback quality, limited student voice in assessment design, and lack of clarity in expectations hindered students’ sense of control and ownership of learning. Conclusion: Assessment practices significantly influence the development of learner agency among medical students. Enhancing the quality of feedback, promoting opportunities for co-agency, and designing student-inclusive assessment systems may strengthen learner empowerment and self-directed learning within medical education.

Medical students frequently experience inconsistent and inadequately structured mentorship. Moreover, research shows that mentorship programmes tend to focus narrowly on academic monitoring without addressing the underlying factors contributing to students' holistic developmental needs. Recognising this gap, the present study seeks to explore the experiences and needs of undergraduate medical students regarding mentorship during their medical training. This study was guided by three theories: Kram's Mentoring Functions Theory, Social Support Theory, and Developmental Network Theory. Nine participants were purposely sampled based on their enrolment in Semesters 4-8. A descriptive qualitative research design with semi-structured individual interviews conducted via Microsoft Teams was employed for data gathering. The interviews were transcribed, and the data were analysed using Braun and Clarke's six steps of thematic analysis. The themes for the mentorship experiences and needs were constructed. The experience themes include "Mentorship as Multifaceted Support System," "Quality of Mentor-Mentee Relationship and Communication," "Mentorship as Catalyst for Growth," "Mentor Attributes and Credibility," and "Structure of Systems." Meanwhile the themes for need of mentorship include "Academic Support," "Trusting Relationship," "Driver of Growth," "Systems that Enable Support," and "Credible, Well-Prepared Mentors." Students provided insights on their mentorship experiences and needs as well as its effects on their personal, academic, and professional development. Several significant trends were found related to academic guidance, emotional support, career preparation, networking, and research engagement. However, students frequently expressed concerns about poor mentor responsiveness, inadequate career guidance, dysfunctional communication infrastructure, and strong emotional impact from a lack of proactive support and unclear expectations. The results supported the need for comprehensive mentorship programmes focusing on career planning, time management, stress management, networking, and research competencies, whilst creating supportive environments characterised by psychological safety, genuine engagement, and structured meeting frequencies rather than perfunctory monitoring. The study's findings add to the literature on mentorship programmes aimed at addressing students' multidimensional needs and promoting holistic wellbeing during medical training. The insights from the undergraduate medical students' viewpoint are helpful for institutions in designing evidence-based mentorship programmes.

Epstein-Barr virus (EBV) is a class I carcinogen human herpes virus which has infected 90% of humanity and most prevalent among Asians, especially Chinese. After primary infection, EBV establishes the lifelong virus carrier state. EBV can be detected in two different tissues namely, B lymphocytes and epithelial cells. EBV is linked to the pathogenesis of a variety of human tumors and disorders, such as Burkitt’s lymphoma, and nasopharyngeal carcinoma. Algae are a potential source of antiviral compounds; however, there have been very few reports on the antiviral activity of microalgae extracts against EBV. The objective of this study was to investigate the antiviral activity of extracts from three microalgae, namely Ankistrodesmus convolutus UMACC 101, Synechococcus elongatus UMACC 105 and Spirulina platensis UMACC 161 against EBV in Burkitt’s lymphoma (BL) cell lines. Three EBV-positive BL cell lines, namely Akata, B95-8 and P3HR-1 were used as in vitro study model. A bioassay-guided fractionation approach was used for the screening of antiviral activity. The antiviral activity of the microalgae extracts was elucidated based on their inhibition efficacy in reducing number of cell-free viral particles being released by chemically induced lytic BL cells. This was assessed by quantifying the cell-free DNA using real-time PCR technique. In addition, the inhibition activity of microalgae extracts against the expression of the viral proteins LMP1, EBNA1 and ZEBRA in BL cells was assessed using immunocytochemistry technique. Two antiviral drugs namely acyclovir and foscarnet were chosen as positive controls. Methanol extracts from Ankistrodesmus convolutus and Synechococcus elongatus displayed low cytotoxicity (IC50 >200 µg/mL) and reduced the cell-free EBV viral load most effectively (EC50 <0.01 µg/mL) and thus, displayed high therapeutic index (>28000). The extracts decreased the expression of EBNA1 (>45%), LMP1 (>38%) and ZEBRA (>67%) effectively in P3HR-1 cells. After column chromatography fractionation, the non-polar fraction of the extract from Synechococcus elongatus (SEF1) reduced the amount of cell-free EBV DNA most effectively (EC50= 2.9μg/mL; therapeutic index >69) with low cytotoxicity (IC50 >200 μg/mL). SEF1 inhibited the expression of EBNA1 and ZEBRA (>40%) effectively in P3HR-1 cells. When SEF1 was further fractionated using HPLC, the sub-fraction SEF1’a was most active in reducing the cell-free EBV DNA (EC50= 1.38µ/mL; therapeutic index >14.5). It inhibited the expression of LMP1 moderately (25%) in P3HR-1 cells. The microalgae extracts did not interact with the cytoskeleton components (actin and tubulin) of BL cells during the release of cell-free EBV particles as revealed by the immunofluorescence study. The active constituents in the microalgae extracts tested might consist of pigments such as chlorophylls, carotenoids, phaeophytins and phycobilins. In conclusion, methanol extracts from Ankistrodesmus convolutus, Synechococcus elongatus and Spirulina platensis showed antiviral activity by inhibiting the release of EBV from the BL cells and the expression of the viral proteins LMP1, EBNA1 and ZEBRA in the host cells. The potential of the microalgae as a source of antiviral drugs against EBV is worth exploring.

Introduction: Currently there is an uptrend in the provision of Extended Pharmacy Services (EPS) among community pharmacists (CPs). EPS referred to various services that beyond the traditional pharmacists’ roles of dispensing medicines such as public health educational program, clinical services and medicine use review. However, such services were only reported in other developed countries but not in Malaysia. Within this context, the present study aimed to explore the types of EPS available, CPs’ perceptions and attitudes towards the provision of EPS. This study also identified the perceived barriers and facilitators towards the provision of EPS. Methods: A descriptive cross-sectional study was conducted. A total of 236 samples were collected through convenience sampling around Selangor and Kuala Lumpur region. The questionnaire consisted of 6 domains including demographic profile, the type of EPS, pharmacist’s perception, perceived barriers, perceived facilitator and pharmacist attitude towards the provision of EPS. All questions were scored using five-point Likert Scale. All the data were analysis through SPSS using descriptive statistics analysis, Mann-Whitney and Kruskal Wallis test. Results: CPs are willing to provide EPS in near future. For the time being, health screening test were the most performed EPS. CPs often/always performed blood pressure test, (97.8%), glucose test (96.6%) and cholesterol test (83.0%). Besides that, the often/always performed counselling sessions for cough and cold (95.8%) and nutritional supplement (94.5%). However, other advanced services like smoking cessation and weight management services were not frequently performed. The top 5 perceived barriers identified includes the lack of standardized practice model for EPS (87.3%), high pressure on generating sales (86.4%), lack of patient awareness (84.4%), 4 lack of access to patient medical record (83.5%) and shortage of time (80.5%). Whereas the support and encouragement from government and other pharmacy organization were identified as the most prominent facilitator towards the implementation of EPS. Conclusion: CPs showed a positive attitude towards the provision of EPS. For the time being, there are various EPS that had already been developed and implemented. However, the identified barriers should be intervened, and facilitators should be executed to implement EPS successfully in near future. Keywords: Pharmaceutical Care Services, Extended Pharmacy Services, Expanded Pharmacy Services, Enhanced Pharmacy Services, Barriers, Facilitators, Community Pharmacy

Dietary fat when consumed reduces hunger and impairs food intake by eliciting satiety signals and these signals are evoked by entry of triacylglycerol after hydrolization to fatty acids into the small intestine. 1,3-dipalmitoyl-2-oleoyglycerol (POP-), 1,3-distearoyl-2-oleoylglycerol (SOS-) and 1,2,3-triolein (OOO-) type of fats have different melting characteristics that may affect postprandial blood lipids, gut hormone concentrations, insulinemic response and selected cardiovascular disease markers in human volunteers. The main objective of this study is to compare the effects of edible fats with either palmitic acid (16:0) (palm mid-fraction) or stearic acid (18:0) (shea stearin) predominantly at the sn-1 and sn-3 positions on postprandial lipemia and gut hormone concentrations. A randomized, double-blind crossover (3 × 3 arms) orthogonal Latin-square design was used on 36 healthy adults (18 males, 18 females; mean age = 23 years). Each subject received 3 different test muffins (each containing 53 g of test fat) in random order separated by 2 weeks over a 6-week period. The test fats of different melting points were palm mid- fraction (PMF; POP-rich), shea stearin (SS; SOS-rich) and high- oleic sunflower oil (HOSF; OOO-rich) During a postprandial test, each subject was provided with a test muffin plus milkshake (total 3.67 MJ or 876 kcal) in the morning and blood samples were collected at half-hourly intervals until 4.0 hours. No significant difference (p>0.05) was observed between the 3 test meals for postprandial responses in plasma TC, Lp(a), apo(B), NEFA, GLP-1, PYY, ghrelin, VAS, PAI-1, IL-6, TNF-α, glucose, insulin and satiety (VAS scores). Plasma TAG peaked at about 4 hours; levels in the PMF- and HOSF- subjects were significantly higher (p<0.05) compared with SS-subjects after 90 minutes. PMF and HOSF exerted a higher postprandial GIP response (p<0.05) as compared to SS. Plasma C-peptide levels, as a measure of insulinemic response, rose sharply 5.5- folds in all groups, peaking after 90 minutes; levels in the SS group declined at a faster rate (p<0.05) than in the PMF- and HOSF- groups. The POP- and OOO- fats induced similar effects on all the biochemical/physiological outcome measures investigated. In contrast, the SOS- type fat (shea stearin) induced a slower rise (p<0.05) in postprandial TAG and GIP levels and a faster return of plasma C-peptide levels to baseline.

Multidrug resistance (MDR) is a serious problem that causes failure in chemotherapy. P-glycoprotein (P-gp) is widely distributed in the intestine, liver and kidneys contributing as one of the major mechanism that causes efflux of chemotherapeutic agents, resulting in ineffective therapeutic level during treatment. However, recent advances in drug delivery systems have made it possible to circumvent MDR issues. In this study, an attempt was made to develop Poly-lactide co-glycolide (PLGA) nanoparticles for oral chemotherapy by co encapsulating doxorubicin (Dox) as the chemotherapeutic agent and cyclosporine-A (CysA) as the chemosensitizer that will function to inhibit P-gp to increase uptake of Dox. High Performance Liquid Chromatography (HPLC) methods were developed and validated for detection and quantification of Dox and CysA. Optimized Dox-CysA nanoparticles were reported to be in 214±4.56 nm of size, zeta potential of -30.1±9.42 and polydispersity index of 0.139±0.09.Transmission Electron Microscopy (TEM) image shows spherical nanoparticles. Differential Scanning Calorimetry (DSC) thermogram indicates Dox and CysA exist in amorphous form in PLGA nanoparticles. Dox and CysA nanoparticles were formulated separately alongside optimized nanoparticle for comparison purposes in drug release and cytotoxicity studies. Dox released slowest from Dox-CysA nanoparticles whilst CysA from CysA nanoparticles in Phosphate Buffer Saline (PBS), promising a controlled release. Cytotoxicity studies conducted on Caco 2 cell line, showed the lowest cell viability of 34%, for Dox-CysA loaded nanoparticles and 76.8% for Dox loaded nanoparticles while a combination of Dox loaded nanoparticles and CysA loaded nanoparticles at the same concentration gave a cell viability of 47.1%, with all treatment containing Dox at 0.71μg/ml. Co-administration of CysA together with Dox in the same nanoparticles formulation proves to increase the uptake of Dox in vitro.