Introduction: p16 gene plays an important role in the normal cell cycle regulation. Methylation of p16 has been reported to be one of the epigenetic events contributing to the pathogenesis of diffuse large B-cell lymphoma (DLBCL) which occurring at varying frequency. DLBCL is an aggressive and high-grade malignancy which accounts for approximately 30% of all non-Hodgkin lymphoma cases. However, little is known regarding the epigenetic alterations of p16 gene in DLBCL cases in Malaysia. Therefore, the objective of this study was to examine the status of p16 methylation in DLBCL. Methods: A total of 88 formalin-fixed paraffin-embedded DLBCL tissues retrieved from two hospitals located in the east coast of Malaysia, namely Hospital Tengku Ampuan Afzan (HTAA) Pahang and Hospital Universiti Sains Malaysia (HUSM) Kelantan, were chosen for this study. DNA specimens were isolated and subsequently subjected to bisulfite treatment prior to methylation specific-PCR. Two pairs of primers were used to amplify methylated and unmethylated regions of p16 gene. The PCR products were then separated using agarose gel electrophoresis and visualised under UV illumination. SPSS version 12.0 was utilised to perform all statistical analysis. Result: p16 methylation was detected in 65 of 88 (74%) samples. There was a significant association between p16 methylation status and patients aged >50 years old (p=0.04). Conclusion: Our study demonstrated that methylation of p16 tumor suppressor gene in our DLBCL cases is common and significantly increased among patients aged 50 years and above. Aging is known to be an important risk factor in the development of cancers and we speculate that this might be due to the increased transformation of malignant cells in aging cell population. However, this has yet to be confirmed with further research and correlate the findings with clinicopathological parameters.

Objective Microplastics (MPs) contamination is a growing emerging concern with potential health risks, particularly for vulnerable populations such as infants. This study aims to develop and optimize a method for isolating MPs from human breast milk, a complex biological matrix with high lipid and protein content that complicates detection. The goal is to evaluate the effectiveness of a combined chemical extraction approach for isolating MPs and identifying their physical characteristics and polymer types. Methods Breast milk samples were ethically obtained from the Halimatussaadia Mother’s Milk Centre (HMMC). Lipids were removed using hexane followed by potassium hydroxide (KOH) and hydrogen peroxide (H₂O₂) treatments to digest organic matter. The chemical extraction process was optimized with varying concentrations: hexane (0.5:1, 1:1), 30% H₂O₂ (1:2, 1:4), and 10% KOH (1:1, 1:3). The treated samples were analysed using Attenuated Total Reflectance Fourier-Transform Infrared (ATR-FTIR) spectroscopy to identify MPs and determine polymer types. Results This study presents an optimized method combining hexane, KOH, and H₂O₂ to isolate microplastics (MPs) from human breast milk, identifying 12 MPs polymer types, including polyethylene (PE), polypropylene (PP), and polyethylene terephthalate (PET). This approach improves detection accuracy and provides a foundation for future research into potential health risks associated with MPs exposure in infants. However, due to technological limitations, the most prevalent MPs polymer in the samples could not be confirmed. Despite this, the study demonstrated the reliability of the combined hexane-KOH –H₂O₂ approach for isolating MPs from human milk. Conclusions This study presents an effective and optimized method for isolating MPs from complex biological fluids such as human breast milk. The findings provide a foundation for future research investigating potential health risks associated with MPs exposure in infants, highlighting the need for larger sample sizes and more advanced analytical techniques.

MGMT and SPOCK2 Promoter Methylation in Diffuse Large B-Cell Lymphoma: A Study in Two Tertiary Health Centres in the East Coast of Malaysia

Background: Milk kinship, in some cultures, posits that infants who receive breast milk from non-biological mothers establish a consanguinity, prohibiting future marriage between the recipient infants and the biological children of the breastfeeding mother. This kinship may be influenced by specific miRNAs in breast milk that modulate gene expression epigenetically, leaving marks within the recipient infants. Research Aim: This study aims to identify lactation-specific miRNAs shared among milk siblings, compare miRNA similarity between a breastfeeding mother’s milk and milk siblings’ plasma, and determine their persistence post-infancy. Methods: RNA was extracted from plasma and breast milk samples, followed by NanoString miRNA analysis to profile individual miRNAs. Differential Gene Expression (DGE) analysis using the R studio NOISeq package identified significantly expressed miRNAs across all samples. Results: Detailed comparisons revealed that several similarly detected miRNAs in milk siblings were present in their corresponding breastfeeding mother’s milk. DGE analysis identified five miRNAs: hsa-let-7b-5p, hsa-miR-223-3p, hsa-miR-873-3p, hsa-miR-23a-3p, and hsa-miR-378e, with hsa-miR-23a-3p exhibiting the most consistent expression pattern throughout the study. Conclusion: This study successfully identifies potential lactation-specific miRNAs that may be shared between milk siblings, providing valuable insights and laying a foundation for future identification of milk kinship biomarkers. However, despite promising findings, current evidence remains insufficient to establish a definitive link between these miRNAs and milk kinship. Further research is required to confirm these findings.

This study successfully developed and evaluated a multi-step digestion protocol for the effective extraction and identification of microplastics (MPs) from human breast milk (HBM). The method combined hexane-mediated lipid removal, enzymatic digestion using Candida rugosa lipase at a milk-to-enzyme ratio of 20:1, protein digestion with potassium hydroxide (KOH) at 1:3 ratio, and oxidative treatment with hydrogen peroxide (H₂O₂) at 10% concentration. Ethanol was found essential for reducing sample viscosity, particularly post-saponification, thereby facilitating efficient filtration. Results showed that extended incubation times during enzymatic digestion notably enhanced lipid removal and improved clarity of the digestate, contributing to higher extraction efficiency. This protocol establishes a reproducible framework for reliable isolation and downstream spectroscopic identification of MPs in breast milk. Despite these advances, improvements in mechanical handling and contamination control are necessary to further enhance accuracy and consistency. The present findings contribute important methodological advancements to the field and support the urgent need for standardised, scalable protocols in monitoring neonatal exposure to environmental microplastic contaminants. Future work should expand on validation across diverse sample sets and explore correlations between MP presence and neonatal health outcomes.